Nsaid formulations, based on highly adaptable aggregates, for improved transport through barriers and topical drug delivery

ABSTRACT

The invention describes novel formulations of nonsteroidal anti-inflammatory drugs (NSAIDS) based on complex aggregates with at least three amphipatic components suspended in a suitable, e.g. pharmaceutically acceptable, polar liquid medium. A suitably ionised NSAID is one of the two, amongst said three, components that tends to destabilise lipid membranes, the other system component with such activity being typically a surfactant. In contrast, the remaining amongst said at least three amphipatic components typically forms a stable lipid membrane on it&#39;s own. An essential characteristics of the resulting, relatively large, aggregates is an improved ability to penetrate pores, in a semi-permeable barrier, at least 30%, and often much smaller than the average diameter of the complex aggregate. This enables said aggregates to mediate NSAID transport through semi-permeable barriers including mammalian skin. As a result of the skin penetration by NSAID loaded large aggregates, the drug delivered transcutaneously with such carriers gets deeper into the tissue than the corresponding NSAID from a solution on the skin surface. This is believed to be due to the special ability of suitable large carriers to bypass the local sink of blood capillaries at the epidermal-dermal junction in the skin. The carrier-mediated delivery of locally applied NSAIDs thus allows therapy of deep tissues under the drug administration site, which is medically highly desirable.

The present application is a continuation of U.S. application Ser. No.10/357,617, filed on Feb. 4, 2003, which claims the benefit of U.S.provisional application No. 60/417,847 filed on Oct. 11, 2002, each ofwhich is incorporated herein by reference in its entirety.

FIELD OF INVENTION

This invention deals with novel formulations of nonsteroidalanti-inflammatory drugs (NSAIDs) based on complex, extended surfaceaggregates comprising at least three amphipatic components. One of thesecomponents is capable of forming stable, large bilayer membranes on it'sown. The other at least two amphipatic components, including an NSAID,tend to destabilise such membranes. Said aggregates are normallysuspended in a suitable, e.g. pharmaceutically acceptable, polar liquidmedium, which also affects NSAID ionisation. The selection of the secondamphipatic membrane destabilising component, which is typically a(co)surfactant, can boost the deformability of the resulting mixedextended surface aggregates. This effect may be supported by judiciouschoice of the other system components. The invention enables animprovement of barrier penetration and drug delivery by such aggregates.The invention also teaches how to select most appropriate NSAIDconcentration, the right total amphipat concentration and, in case,amphipat ionisation in the resulting mixed aggregate suspension. Theinvention further relates to preparation and application of theresulting suspension in pharmaceutical formulations, with a focus onepicutaneous application on, or less frequently in, warm bloodedcreatures.

BACKGROUND INFORMATION

The current state of the art in NSAID delivery through the skin istransdermal drug diffusion, which is proportional to the drugconcentration on the skin and inversely proportional to the skin barrierresistance, which is tantamount to saying that diffusion is proportionalto the skin permeability.

Solubility of typical NSAIDs is in the range 1 μg/ml to between 0.5mg/ml and 10 mg/ml for the pH range between 1 and 7.5. This correspondsto a few μM and up to a few tens of mM, high values being alwaysmeasured in least acidic solutions (pH>>pK_(a)) where NSAIDs are partlyor completely ionised, the solubility at pH<<pK_(a) always being verylow. To maximise diffusive NSAID transport through the skin one shouldtherefore always use the highest tolerable pH, which can exceed thevalue of 9.

Taken the limitations of maximum NSAID solubility, attempts have beenmade to improve NSAID permeation (diffusion) through the skin by usingpermeability or permeation enhancers. Permeability enhancers increaseNSAID flux through the barrier for a given drug concentration, but donot much affect the depth of drug distribution. Further, use ofconventional lipid formulations on the skin does not affect thislimitation.

For example, Henmi et al. 1994 (Chem Pharm Bull 42:651-655) used threedifferent NSAIDs (ketoprofen, flurbiprofen and ibuprofen) in an oilygel, formed by hydrogenated soybean phospholipids (which forms verystiff membranes) and applied the preparation on the skin. The conclusionwas that such lipids have no permeation enhancing effect for the skinbut rather solubilise the test drug.

Burnham et al. 1998 (Clin J Sport Med 8:78-81) used a block co-polymerof polyethylene and an unspecified polypropylene glycol (pluronic),which generally is a poor membrane destabilising amphipat, to apply anNSAID on the skin. An unspecified lecithin based liposomal organo-gel(PLO) was furthermore used three times daily for one week, followed by aweekly “washout” period without using the gel. The authors noted thatonly a thin tissue layer under the skin was treated, thus implying thatany apparently positive result could be due to free drug diffusion fromPLO through the skin. Organo-gel consequently has served as merely asuperficial reservoir.

Vyas et al. (J Microencapsul 12:149-54, 1995) incorporated diclofenacinto multilamellar, 1-5 μm large liposomes at pH=7.4 that were appliedon the skin under different conditions. The resulting systemic drugavailability was then studied. The resulting mixed lipid vesicles wereincorporated in an ointment base and were applied on the skin of rats.However, skin poration by ultrasound was required to achieve anysubstantial transdermal delivery of the drug, and most of the testedNSAID was typically found at the site of application.

Schramlova et al. (Folia Biol (Praha) 43:195-199, 1997) associatedibuprofen with liposomes prepared from soybean phospholipid supplementedwith 10 rel-% cholesterol, the knowledge in the art being that thelatter is a membrane stiffening agent. The formulation with a pH=7.4 wasinjected intramuscularly or applied under occlusion on the skin. NSAIDfrom lipid vesicles occasionally decreased the rat leg edema slightly,but not significantly, better than the drug from a conventional creambut less than an NSAID injection. This paper therefore teaches the useof a membrane stabilising component (cholesterol) rather than of amembrane destabilising component.

Saunders et al. (J Pharm Pharm Sci 2:99-107, 1999), studying the skinpermeation enhancement, also used liposomal structures of unspecifiedcomposition and morphology, which were claimed to be present in the MZLlotion and in a comparator gel (both prepared by Meyer Zall Laboratories(MZL)), and loaded with sodium diclofenac. The presence of oil in theoil/water base in the MZL formulation, which diminishes lipid aggregatedeformability, and occludes the skin, if nothing else precludedefficient drug delivery by vesicle through the skin.

Calpena et al. (Arzneimittelforschung 49:1012-1017, 1999) studieddiclofenac permeation through human skin from 6 semisolid formulationscontaining 1% drug in a complex mixture of gel-forming materialscombined with lecithin (2.5% of unspecified quality) and cholesterol(0.5%). However, the results of the studies suggest that use of lipidvesicles is not beneficial (Calpena et al., 1999).

Skin permeability data for ibuprofen lysinate was studied, showingpractically equal permeability rates for the drug in solution or inmixed micelles (containing soy-bean phosphatidylcholine) and nearly3-times lower rate for the corresponding liposomal dispersion (Stoye etal., 1998 (Eur J Pharm Biopharm 46:191-200). Liposomes therefore wereconcluded to be useless in terms of supporting transdermal drugtransport in the described system.

SUMMARY OF THE INVENTION

We have found, unexpectedly, that various combinations of at least twoamphipatic components one of which is an NSAID, which can substantiallydestabilise a lipid-based, otherwise stable extended surface aggregate,typically in the form of a bilayer membrane, can synergisticallyincrease the resulting at least three-component aggregate adaptability.In parallel, the aggregate (membrane) shape deformability issynergistically augmented. Consequently, the flux of such aggregatesuspension through narrow pores is increased and/or the characteristicpressure that drives certain flux through the corresponding porousbarrier is lowered.

The capability of said at least three-component aggregates to movethrough a semi-permeable barrier is thus facilitated. This finding issurprising given that the droplets covered by a bi-component bilayermembrane already have an appreciable barrier crossing capabilitycompared to droplets enclosed by a simple lipid bilayer.

The increase of adaptability of said extended surface aggregates with atleast three amphipatic components and/or the lowering of the pressurethat is needed to make such aggregates move through a biological barrierhas important, and unexpected, practical consequences. Specifically,when said aggregates are applied on the skin, as an example for abiological semi-permeable barrier, the transport of the aggregateassociated NSAIDs through such barrier is increased and reaches further.The latter observation is explicable in terms of differential clearancein the superficial skin layers, where cutaneous blood drainage resides,of the drug, which can enter directly into blood capillaries, and ofdrug-loaded aggregates, which are too big to enter such capillaries.This means that NSAID carriers move further than the drug from solution,allowing deeper tissues to be treated with NSAIDs under the drugapplication site on the skin. Convincing evidence for this is given inone of Practical Examples. Such finding is not expected taken thatsimple NSAID-phospholipid combinations already ensure better and deeperdrug transport through the skin than conventional preparations based onNSAID solutions. In the present invention, the general terms employedhereinbefore and hereinafter have the following meanings.

The term “aggregate” denotes a group of more than just a few amphipatsof similar or different kind. Typically, an aggregate referred to inthis invention contains at least 100 molecules, i.e. has an aggregationnumber n_(a)>100. More often aggregation number is n_(a)>1000 and mostpreferably n_(a)>10.000. An aggregate comprising an aqueous coresurrounded with at least one lipid (bilayer) membrane is called a lipidvesicle, and often a liposome.

The term aggregate “adaptability” is defined in this document as theability of a given aggregate to change easily, and more or lessreversibly, its properties, such as shape, elongation ratio, and surfaceto volume ratio. Adaptability also implies that an aggregate can sustainunidirectional force or stress, such as a hydrostatic pressure, withoutsignificant fragmentation, as is defined for the “stable” aggregates. Aneasy and reversible change in aggregate shape furthermore implies highaggregate deformability and requires large surface-to-volume ratioadaptation. For vesicular aggregates, the latter is associated withmaterial exchange between the outer and inner vesicle volume, i.e. withat least transient vesicle membrane permeabilisation. The experimentallydetermined capability of given aggregate suspension to pass throughnarrow pores in a semi-permeable barrier thus offers simple means forfunctionally testing aggregate adaptability and deformability (videsupra), as is described in the Practical Examples.

To assess aggregate adaptability it is useful to employ the followingmethod:

-   1) measure the flux j_(a) of aggregate suspension through a    semi-permeable barrier (e.g. gravimetrically) for different    transport-driving trans-barrier pressures delta p;-   2) calculate the pressure dependence of barrier penetrability P for    the given suspension by dividing each measured flux value with the    corresponding driving pressure value: P (delta p)=j_(a) (delta    p)/delta p;-   3) monitor the ratio of final and starting vesicle diameter 2r_(ves)    (delta p)/2r_(ves,0) (e.g. with the dynamic light scattering),    wherein 2r_(ves) (delta p)/is the vesicle diameter after    semi-permeable barrier passage driven by delta p and 2r_(ves,0) is    the starting vesicle diameter, and if necessary making corrections    for the flow-rate effects;-   4) align both data sets P (delta p) vs. r_(ves) (delta p)/r_(ves,0),    to determine the co-existence range for high aggregate adaptability    and stability; it is also useful, but not absolutely essential, to    parameterise experimental penetrability data within the framework of    Maxwell-approximation in terms of the necessary pressure value p*    and of maximum penetrability value P_(max), which are defined    graphically in the following illustrative schemes.

It is plausible to sum-up all the contributions to a moving aggregateenergy (deformation energy/ies, thermal energy, the shearing work, etc.)into a single, total energy. The equilibrium population density ofaggregate's energetic levels then may be taken to correspond toMaxwell's distribution. All aggregates with a total energy greater thanthe activation energy, E f E_(A), are finally concluded to penetrate thebarrier. The pore-crossing probability for such aggregates is then givenby:

${P(e)} = {1 - {{erf}\left( \sqrt{\frac{1}{e}} \right)} + {\sqrt{\frac{4}{\pi \; e}} \cdot {\exp \left\lbrack {- \frac{1}{e}} \right\rbrack}}}$

e being dimensionless aggregate energy in units of the activation energyE_(A).

It is therefore plausible to write barrier penetrability to a givensuspension as a function of transport driving pressure (=drivingpressure difference) p (=delta p) as:

$\begin{matrix}{{P(p)} = {p_{\max} \cdot \left\{ {1 - {{erf}\left( \sqrt{\frac{p^{*}}{p}} \right)} + {\sqrt{\frac{4p^{*}}{\pi \; p}} \cdot {\exp \left\lbrack {- \frac{p^{*}}{p}} \right\rbrack}}} \right\}}} & \left( {}^{*} \right)\end{matrix}$

P_(max) is the maximum possible penetrability of a given barrier. (Forthe aggregates with zero transport resistance this penetrability isidentical to the penetrability of the suspending medium flux.) p* is anadjustable parameter that describes the pressure sensitivity, and thusthe transport resistance, of the tested system. (For barriers with afixed pore radius this sensitivity is a function of aggregate propertiessolely. For non-interacting particles the sensitivity is dominated byaggregate adaptability, allowing to make the assumption: a_(a)proportional to 1/p*.)

The formula (*), is used in various Practical Examples to calculateaggregate adaptability from suspension flux, or more precisely from thecorresponding penetrability (═P(p)=Flux/Pressure=Flux/p data).

This formula is explained, in more detail, in our copending U.S.application entitled “Aggregates with increased deformability,comprising at least three amphipats, for improved transport throughsemi-permeable barriers and for the non-invasive drug application invivo, especially through the skin”, filed concurrently, the disclosureof which is incorporated herein by reference.

The term “apparent dissociation constant” (“pKa”) refers to the measureddissociation (i.e. ionisation) constant of a drug. This constant formany drugs, including NSAIDs, is different in the bulk and in the homo-or heteroaggregates. For ketoprofen, the pKa in the bulk is approx. 4.4whereas the pKa value measured above the drug association concentrationis approx. 5, and decreases approximately linearly with the inverseionic strength of the bulk solution. pKa of ketoprofen bound to lipidbilayers increases with total lipid concentration as well, and isapprox. 6 and 6.45 in suspensions with 5 w-% and 16 w-% total lipid in a50 mM monovalent buffer, respectively. For diclofenac, the pKa in thebulk is around 4, whereas for this drug in lipid bilayers pKa˜6.1 wasdetermined. The bulk pKa reported in the literature for meloxicam,piroxicam, naproxen, indomethacin and ibuprofen is 4.2 (and 1.9), 5.3,4.2-4.7, 4.5, and 4.3 (or in some reports 5.3), respectively.

The term aggregate “deformability” is closely related to the term“adaptability”. Any major change in aggregate shape that does not resultin a significant aggregate fragmentation is indicative of sufficientaggregate deformability, and also implies a large change in the deformedaggregate surface-to-volume ratio. Deformability can therefore bemeasured in the same kind of experiments as is proposed for determiningaggregate adaptability, or else can be assessed by optical measurementsthat reveal reversible shape changes.

The term “long” used in connection with a fatty residue attached to alipid, a surfactant or a drug implies the presence of 10 to 24 carbonatoms in alkyl, alkenyl, alkoxy, alkenyloxy, or acyloxy chains, whichindividually or together, as the case may be, bear the class name of“fatty chains”. Implicitly included in this term, but not furtherspecified in detail, are “fatty chains” with at least one branched or acyclic, but unpolar or little polar segment.

The term “narrow” used in connection with a pore implies that the porediameter is significantly, typically at least 30%, smaller than thediameter of the entity tested with regard to its ability to cross thepore.

The term “NSAID” (nonsteroidal anti-inflammatory drug) typicallyindicates a chemical entity which acts as lipoxygenase, cyclooxygenase-1or cyclooxygenase-2 antagonist.

Examples include analgesics, for example, salts of substitutedphenylacetic acids or 2-phenylpropionic acids, such as alclofenac,ibufenac, ibuprofen, clindanac, fenclorac, ketoprofen, fenoprofen,indoprofen, fenclofenac, diclofenac, flurbiprofen, pirprofen, naproxen,benoxaprofen, carprofen or cicloprofen; analgesically activeheteroarylacetic acids or 2-heteroarylpropionic acids having a2-indol-3-yl or pyrrol-2-yl radical, for example indomethacin,oxmetacin, intrazol, acemetazin, cinmetacin, zomepirac, tolmetin,colpirac or tiaprofenic acid; analgesically active indenylacetic acids,for example sulindac; analgesically active heteroaryloxyacetic acids,for example benzadac; NSAIDS from oxicame family include piroxicam,droxicam, meloxicam, tenoxicam; further interesting drugs from NSAIDclass are, meclofenamate, and the like.

A list of commonly used NSAIDs is given in the following table:

NSAID Some common trade names Acetaminofene Tylenol Cimicifuga ArtolCholine salicylate-Mg salicylate Trilisate Diclofenac as Na salt:Apo-Diclo, Apo-Diclo SR, Arthrotec, Diclofenac Ect, Novo-Difenac,Novo-Difenac SR, Nu-Diclo, Taro-Diclofenac, Voltaren, Voltaren SR; as Ksalt: Voltaren Rapide Diflunisal Apo-Diflunisal, Dolobid,Novo-Diflunisal, Nu-Diflunisal Etodolac Ultradol Fenoprofen calciumNalfon Floctafenine Idarac Flurbiprofen Ansaid, Apo-Flurbiprofen FC,Froben, Froben SR, Novo-Flurprofen, Nu-Flurbiprofen IbuprofenActiprofen, Advil, Advil Cold & Sinus, Amersol, Apo- Ibuprofen, ExcedrinIB, Medipren, Motrin, Motrin IB, Novo-Profen, Nuprin, Nu-IbufrofenIndomethacin Apo-Indomethacin, Indocid, Indocid SR, Indolec,Novo-Methacin, Nu-Indo, Pro-Indo, Rhodacine Ketoprofen Apo-Keto,Apo-Keto-E, Novo-Keto, Novo-Keto-Ec, Nu-Ketoprofen, Nu-Ketoprofen-E,Orudis, Orudis E, Orudis SR, Oruvail, PMS-Ketoprofen, PMS- Ketoprofen-E,Rhodis, Rhodis-EC Ketorolac tromethamine Acular, Toradol Magnesiumsalicylate Back-Ese-M, Doan's Backache Pills, Herbogesic Mefenamic acidPonstan Nabumetone Relafen Naproxen Apo-Naproxen, Naprosyn, aprosyn-E,Naxen, Novo- Naprox, Nu-Naprox, PMS-Naproxen; or in the sodium form:Anaprox, Anaprox DS, Apo-Napro-Na, Naproxim-Na, Novo-Naprox Sodium,Synflex, Synflex DS Oxyphenbutazone Oxybutazone Phenylbutazone AlkaPhenyl, Alka Phenylbutazone, Apo- Phenylbutazone Butazolidin,Novo-Butazone, Phenylone Plus Piroxicam Apo-Piroxicam, Feldene,Kenral-Piroxicam, Novo- Pirocam, Nu-Pirox, PMS-Piroxicam, Pro-Piroxicam,Rho- Piroxicam Salsalate Disalcid Sodium salicylate Apo-Sulin, Dodd's,Dodd's Extra-Strength, Sulindac, Clinoril, Novo-Sundac, Nu-Sulindac,Sulindac Tenoxicam Mobilflex Tiaprofenic acid Albert Tiafen,Apo-Tiaprofenic, Surgam, Surgam SR Tolmetin sodium Novo-Tolmetin,Tolectin

The term “phospholipid” has, for example, the formula

in which one of the radicals R1 and R2 represents hydrogen, hydroxy orC1-C4-alkyl, and the other radical represents a long fatty chain,especially an alkyl, alkenyl, alkoxy, akenyloxy or acyloxy, each havingfrom 10 to 24 carbon atoms, or both radicals R1 and R2 represent a longfatty chain, especially an alkyl, alkenyl, alkoxy, alkenyloxy or acyloxyeach having from 10 to 24 carbon atoms, R3 represents hydrogen orC1-C4-alkyl, and R4 represents hydrogen, optionally substitutedC1-C7-alkyl or a carbohydrate radical having from 5 to 12 carbon atomsor, if both radicals R1 and R2 represent hydrogen or hydroxy, R4represents a steroid radical, or is a salt thereof. The radicals R1, R2,R3, and R4 are typically selected so as to ensure that lipid bilayermembrane is in the fluid lamellar phase during practical application andis a good match to the drug of choice.

In a phospholipid of the formula 1, R1, R2 or R3 having the meaningC1-C4-alkyl is preferably methyl, but may also be ethyl, n-propyl, orn-butyl.

The terms alkyl, alkenyl, alkoxy, akenyloxy or acyloxy have their usualmeaning. The long fatty chains attached to a phospholipid can also besubstituted in any of usual ways.

A steroid radical R4 is, for example, a sterol radical that isesterified by the phosphatidyl group by way of the hydroxy group locatedin the 3-position of the steroid nucleus.

If R4 represents a steroid radical, R1 and R2 are preferably hydroxy andR3 is hydrogen.

Phospholipids of the formula 1 can be in the form of free acids or inthe form of salts. Salts are formed by reaction of the free acid of theformula II with a base, for example a dilute, aqueous solution of alkalimetal hydroxide, for example lithium, sodium or potassium hydroxide,magnesium or calcium hydroxide, a dilute aqueous ammonia solution or anaqueous solution of an amine, for example a mono-, di- or tri-loweralkylamine, for example ethyl-, diethyl- or triethyl-amine,2-hydroxyethyl-tri-CI-C4-alkyl-amine, for example choline, and a basicamino acid, for example lysine or arginine.

A phospholipid of the formula 1 has especially two acyloxy radicals R1and R2, for example alkanoyloxy or alkenoyloxy, for example lauroyloxy,myristoyloxy, palmitoyloxy, stearoyloxy, arachinoyloxy, oleoyloxy,linoyloxy or linoleoyloxy, and is, for example, natural lecithin(R3=hydrogen, R4=2-trimethylammonium ethyl) or cephalin (R3=hydrogen,R4=2-ammonium ethyl) having different acyloxy radicals R1 and R2, forexample egg lecithin or egg cephalin or lecithin or cephalin from soyabeans, synthetic lecithin or cephalin having different or identicalacyloxy radicals R1 and R2, for example 1-palmitoyl-2-oleoyl lecithin orcephalin or dipalmitoyl, distearoyl, diarachinoyl, dioleoyl, dilinoyl ordilinoleoyl lecithin or cephalin, natural phosphatidyl serine(R3=hydrogen, R4=2-amino-2-carboxyethyl) having different acyloxyradicals R1 and R2, for example phosphatidyl serine from bovine brain,synthetic phosphatidylserine having different or identical acyloxyradicals R1 and R2, for example dioleoyl-, dimyristoyl- ordipalmitoyl-phosphatidyl serine, or natural phosphatidic acid (R3 andR4=hydrogen) having different acyloxy radicals R1 and R2.

A phospholipid of the formula 1 is also a phospholipid in which R1 andR2 represent two identical alkoxy radicals, for example n-tetradecyloxyor n-hexadecyloxy (synthetic ditetradecyl or dihexadecyl lecithin orcephalin), R1 represents alkenyl and R2 represents acyloxy, for examplemyristoyloxy or palmitoyloxy (plasmalogen, R3=hydrogen,R4=2-trimethylammonium ethyl), R1 represents acyloxy and R2 representshydroxy (natural or synthetic lysolecithin or lysocephalin, for example1-myristoyl or 1-palmitoyl-lyso-lecithin or -cephalin; natural orsynthetic lysophosphatidyl serine, R3=hydrogen,R4=2-amino-2-carboxyethyl, for example lysophosphatidyl serine frombovine brain or 1-myristoyl- or 1-palmitoyl-lysophosphatidyl serine,synthetic lysophosphatidyl glycerine, R3=hydrogen, R4=CH₂0H—CHOH—CH₂—,natural or synthetic lysophosphatidic acid, R3=hydrogen, R4=hydrogen,for example egg lysophosphatidic acid or 1-lauroyl-, 1-myristoyl- or1-palmitoyl-lysophosphatidic acid).

The term “semipermeable” used in connection with a barrier implies thata solution can cross transbarrier openings whereas a suspension ofnon-adaptable aggregates 150-200% larger than the diameter of suchopenings cannot achieve this. Conventional lipid vesicles (liposomes)made from any common phospholipid in the gel lamellar phase or else fromany biological phosphatidylcholine/cholesterol 1/1 mol/mol mixture orelse comparably large oil droplets, all having the specified relativediameter, are three examples for such non-adaptable aggregates.

The term sufficiently “stable” means that the tested aggregate does notchange its diameter spontaneously or under reasonable mechanical stress(e.g. during passage through a semipermeable barrier) to a practically,most often pharmaceutically, unacceptable degree. A 20-40% change isconsidered acceptable; the halving of aggregate diameter or a 100%diameter increase is not.

The term “sterol radical” means, for example, the lanosterol,sitosterol, coprostanol, cholestanol, glycocholic acid, ergosterol orstigmasterol radical, is preferably the cholesterol radical, but canalso be any other sterol radical known in the art.

The term “surfactant” also has its usual meaning. A long list ofrelevant surfactants and surfactant related definitions is given in EP 0475 160 and U.S. Pat. No. 6,165,500 which are herewith explicitlyincluded by reference and in appropriate surfactant or pharmaceuticalHandbooks, such as Handbook of Industrial Surfactants or USPharmacopoeia, Pharm. Eu. The following list therefore only offers aselection, which is by no means complete or exclusive, of severalsurfactant classes that are particularly common or useful in conjunctionwith present patent application. This includes ionised long-chain fattyacids or long chain fatty alcohols, long chain fatty ammonium salts,such as alkyl- or alkenoyl-trimethyl-, -dimethyl- and -methyl-ammoniumsalts, alkyl- or alkenoyl-sulphate salts, long fatty chaindimethyl-aminoxides, such as alkyl- or alkenoyl-dimethyl-aminoxides,long fatty chain, for example alkanoyl, dimethyl-aminoxides andespecially dodecyl dimethyl-aminoxide, long fatty chain, for examplealkyl-N-methylglucamides and alkanoyl-N-methylglucamides, such asMEGA-8, MEGA-9 and MEGA-10, N-long fatty chain-N,N-dimethylglycines, forexample N-alkyl-N,N-dimethylglycines, 3-(long fattychain-dimethylammonio)-alkanesulphonates, for example3-(acyldimethylammonio)-alkanesulphonates, long fatty chain derivativesof sulphosuccinate salts, such as bis(2-ethylalkyl)sulphosuccinatesalts, long fatty chain-sulphobetaines, for example acyl-sulphobetaines,long fatty chain betaines, such as EMPIGEN BB or ZWITTERGENT-3-16,-3-14, -3-12, -3-10, or -3-8, or polyethylen-glyco-acylphenyl ethers,especially nonaethylen-glycol-octylphenyl ether, polyethylene-long fattychain-ethers, especially polyethylene-acyl ethers, such asnonaethylen-decyl ether, nonaethylen-dodecyl ether oroctaethylene-dodecyl ether, polyethyleneglycol-isoacyl ethers, such asoctaethyleneglycol-isotridecyl ether, polyethyleneglycol-sorbitane-longfatty chain esters, for example polyethyleneglycol-sorbitane-acyl estersand especially polyethylenglykol-monolaurate (e.g. Tween 20),polyethylenglykol-sorbitan-monooleate (e.g. Tween 80),polyethylenglykol-sorbitan-monolauroleylate,polyethylenglykol-sorbitan-monopetroselinate,polyethylenglykol-sorbitan-monoelaidate,polyethylenglykol-sorbitan-myristoleylatepolyethylenglykol-sorbitan-palmitoleinylate,polyethylenglykol-sorbitan-petroselinylate, polyhydroxyethylene-longfatty chain ethers, for example polyhydroxyethylene-acyl ethers, such aspolyhydroxyethylene-lauryl ethers, polyhydroxyethylene-myristoyl ethers,polyhydroxyethylene-cetylstearyl, polyhydroxyethylene-palmityl ethers,polyhydroxyethylene-oleoyl ethers, polyhydroxyethylene-palmitoleoylethers, polyhydroxyethylene-linoleyl, polyhydroxyethylen-4, or 6, or 8,or 10, or 12-lauryl, miristoyl, palmitoyl, palmitoleyl, oleoyl orlinoeyl ethers (Brij series), or in the corresponding esters,polyhydroxyethylen-laurate, -myristate, -palmitate, -stearate or-oleate, especially polyhydroxyethylen-8-stearate (Myrj 45) andpolyhydroxyethylen-8-oleate, polyethoxylated castor oil 40 (CremophorEL), sorbitane-mono long fatty chain, for example alkylate (Arlacel orSpan series), especially as sorbitane-monolaurate (Arlacel 20, Span 20),long fatty chain, for example acyl-N-methylglucamides,alkanoyl-N-methylglucamides, especially decanoyl-N-methylglucamide,dodecanoyl-N-methylglucamide, long fatty chain sulphates, for examplealkyl-sulphates, alkyl sulphate salts, such as lauryl-sulphate (SDS),oleoyl-sulphate; long fatty chain thioglucosides, such asalkylthioglucosides and especially heptyl-, octyl andnonyl-beta-D-thioglucopyranoside; long fatty chain derivatives ofvarious carbohydrates, such as pentoses, hexoses and disaccharides,especially alkyl-glucosides and maltosides, such as hexyl-, heptyl-,octyl-, nonyl and decyl-beta-D-glucopyranoside or D-maltopyranoside;further a salt, especially a sodium salt, of cholate, deoxycholate,glycocholate, glycodeoxycholate, taurodeoxycholate, taurocholate, afatty acid salt, especially oleate, elaidate, linoleate, laurate, ormyristate, most often in sodium form, lysophospholipids,n-octadecylene-glycerophosphatidic acid,octadecylene-phosphorylglycerol, octadecylene-phosphorylserine, n-longfatty chain-glycero-phosphatidic acids, such asn-acyl-glycero-phosphatidic acids, especially laurylglycero-phosphatidic acids, oleoyl-glycero-phosphatidic acid, n-longfatty chain-phosphorylglycerol, such as n-acyl-phosphorylglycerol,especially lauryl-, myristoyl-, oleoyl- orpalmitoeloyl-phosphorylglycerol, n-long fatty chain-phosphorylserine,such as n-acyl-phosphorylserine, especially lauryl-, myristoyl-, oleoyl-or palmitoeloyl-phosphorylserine, n-tetradecyl-glycero-phosphatidicacid, n-tetradecyl-phosphorylglycerol, n-tetradecyl-phosphorylserine,corresponding-, elaidoyl-, vaccenyl-lysophospholipids, correspondingshort-chain phospholipids, as well as all surface active and thusmembrane destabilising polypeptides. Surfactant chains are typicallychosen to be in a fluid state or at least to be compatible with themaintenance of fluid-chain state in carrier aggregates.

The term “surfactant like phospholipid” means a phospholipid withsolubility, and other relevant properties, similar to those of thecorresponding surfactants mentioned in this application, especially inthe claims 10 and 11. A non-ionic surfactant like phospholipid thereforeshould have water solubility, and ideally also water diffusion/exchangerates, etc., similar to those of a relevant non-ionic surfactant.

Quite detailed recommendations on the preparation of said combinationsis given in EP 0 475 160 and U.S. Pat. No. 6,165,500, which are herewithincluded by reference, using filtering material with pore diametersbetween 0.01 μm and 0.1 μm, more preferably with pore diameters between0.02 μm and 0.3 μm and even more advisable filters with pore diametersbetween 0.05 μm and 0.15 μm to homogenise final vesicle suspension, whenfiltration is used for the purpose. Other methods of mechanicalhomogenisation or for lipid vesicle preparation known in the art areuseful as well.

The lipids and certain surfactants mentioned hereinbefore andhereinafter having a chiral carbon atom can be present both in the formof racemic mixtures and in the form of optically pure enantiomers in thepharmaceutical compositions that can be prepared and used according tothe invention.

To manufacture a pharmaceutical formulation, it may advisable ornecessary to prepare the product in several steps, changing temperature,pH, ion strength, individual component (e.g. membrane destabiliser,formulation stabiliser or microbicide) or total lipid concentration, orsuspension viscosity during the process.

A list of relevant and practically useful thickening agents is givene.g. in PCT/EP98/08421, which also suggests numerous interestingmicrobicides and antioxidants; the corresponding sections ofPCT/EP98/08421 are therefore included into the present application byreference. Practical experiments have confirmed that sulphites, such assodium sulphite, potassium sulphite, bisulphite and metasulphite; andpotentially other water soluble antioxidants, which also contain asulphur or else a phosphorus atom (e.g. in pyrosulphate, pyrophosphate,polyphosphate, erythorbate, tartrate, glutamate, and the like or evenL-tryptophan, ideally with a spectrum of activity similar to that ofsulphites) offer some anti-oxidative protection to said formulations,final selection being subject to regulatory constraints. Any hydrophilicantioxidant should always be combined with a lipophilic antioxidant,however, such as BHT (butylated hydroxytoluene) or BHA (butylatedhydroxyanisole).

In one important aspect of the present invention, the invention providespreparations, based on a suspension of extended surface aggregates in aliquid medium comprising at least one first amphipatic component; atleast one second amphipatic component; at least one third amphipaticcomponent, the first amphipatic component being a vesicle membraneforming lipid component, the second and third component being membranedestabilising components, wherein the third component is a non-steroidalanti-inflammatory drug (NSAID) such that said aggregates are capable ofpenetrating semi-permeable barriers with pores at least 50% smaller thanthe average aggregate diameter before the penetration without changingtheir diameter by more than 25%.

It is another aspect of the invention suspensions of extended surfaceaggregates in a liquid medium are provided, comprising: at least onefirst amphipatic component; at least one second amphipatic component; atleast one third amphipatic component; the first amphipatic componentbeing an aggregate, typically a membrane, forming lipid component; thesecond and third component being aggregate, typically membrane,destabilising components; wherein the third component is a NSAID, suchthat the extended surfaces formed by the first and second componentalone or else by the first and third component alone, the second orthird component, respectively, being present at a relative concentrationX, have a lower propensity to overcome barriers with pores having adiameter at least 50% smaller than the average aggregate diameter,before the pore crossing, than the extended surfaces formed by thefirst, second and third component together, if the second and thirdcomponents are present at or below the combined relative concentrationof X. More specifically, this e.g. means that: a) said extended surfacesformed by the first and second component alone, the second componentbeing present at a relative concentration X, have a lower propensity toovercome barriers with the pores at least 50% smaller than the averageaggregate diameter before the pore crossing than the extended surfacesformed by the first, second and third component, if the second and thirdcomponents are present at or below a combined concentration of X; orelse b) such extended surfaces formed by the first and third componentalone, the third component being present at a relative concentration X,have a lower propensity to overcome barriers with the pores at least 50%smaller than the average aggregate diameter before the pore crossingthan extended surfaces formed by the first, second and third component,the second and third components together being present at or below aconcentration of X.

In yet another aspect of the invention extended surface aggregatessuspended in a liquid medium are provided, comprising: at least onefirst amphipatic component; at least one second amphipatic component; atleast one third amphipatic component; the first amphipatic componentbeing a membrane forming lipid component; the second and third componentbeing membrane destabilising components, such that the third componentis a NSAID; and the inclusion of the second or third component to anotherwise two amphipatic-component mixture increases the suspension fluxthrough the pores at least 50% smaller than the average aggregatediameter before the penetration in comparison with the flux of thesuspension containing aggregates comprising merely the first and secondor the first and third components, respectively. More specifically, theinclusion of the third component increases the flux of said suspensioncompared with the flux of the suspension containing simpler aggregatescomprising merely the first and second component or else the inclusionof the second component increases the flux of said suspension comparedwith the flux of the suspension containing simpler aggregates comprisingmerely the first and third component.

In a further aspect of this invention extended surface aggregatessuspended in a liquid medium comprise: at least one first amphipaticcomponent; at least one second amphipatic component; at least one thirdamphipatic component; the first amphipatic component being a membraneforming lipid component; the second and third component being membranedestabilising components, such that the third component is a NSAID andthat the addition of the second or third component to an originally twocomponent mixture increases aggregate adaptability of the resultingextended surface aggregates with at least three components compared tothe aggregates containing respective combinations of the first and thethird or the first and the second components alone. More specifically,the inclusion of the third component increases the aggregateadaptability of an extended surface aggregate comprising the first andsecond components alone; or else, the inclusion of the second componentincreases the aggregate adaptability of an extended surface aggregatecomprising the first and third components alone.

Yet another aspect of this invention provides extended surfaceaggregates suspended in a liquid medium, comprising: at least one firstamphipatic component; at least one second amphipatic component; at leastone third amphipatic component; the first amphipatic component being amembrane forming lipid component; the second and third component beingaggregate destabilising components, such that the third component is anNSAID; and the inclusion of the second or third component to anotherwise two amphipatic component mixture lowers the driving pressurerequired for aggregate penetration of pores at least 50% smaller thanthe average aggregate diameter before the penetration in comparison withthe aggregates comprising merely the first and second or the first andthird components, respectively. More specifically, the inclusion of thesecond component lowers the driving pressure required for aggregatepenetration of pores at least 50% smaller than the average aggregatediameter before the penetration in comparison with the aggregatescomprising merely the first and third components; alternatively, theinclusion of the third component lowers the driving pressure requiredfor aggregate penetration of pores at least 50% smaller than the averageaggregate diameter before the penetration in comparison with theaggregates comprising merely the first and second components.

It is a further aspect of this invention to provide extended surfaceaggregates suspended in a liquid medium, comprising: at least one firstamphipatic component; at least one second amphipatic component; at leastone third amphipatic component; the first amphipatic component being amembrane forming lipid component; the second and third component beingmembrane destabilising components, such that the third component is anNSAID and the inclusion of the second or third component to an otherwisetwo amphipatic component mixture increases the deformability of extendedsurface aggregates compared with the aggregates comprising merely thefirst and second or the first and third component, respectively. Morespecifically, the inclusion of the third component increases thedeformability of the extended surface aggregates compared with theaggregates comprising merely the first and second component;alternatively, the inclusion of the second component increases thedeformability of the extended surface aggregate compared with theaggregates comprising merely the first and third component.

The invention teaches preparation and use of said extended surfaceaggregates in the form of membrane-enclosed, liquid-filled vesicles,whereby said first component is a membrane-forming lipid, and saidsecond and third components are membrane-destabilising components.

The invention includes suspensions of extended surface aggregates in aliquid medium comprising: at least one first amphipatic component; atleast one second amphipatic component; at least one third amphipaticcomponent; the first amphipatic component being a membrane forming lipidcomponent; the second and third component being membrane destabilisingcomponents, such that the third component is a non-steroidalanti-inflammatory drug (NSAID) and such that said extended surfaceaggregates can penetrate intact mammalian skin, thus increasing NSAIDconcentration in the skin and/or increasing the reach of NSAIDdistribution below the skin, in comparison with the result of the sameNSAID application in a solution on the skin. In a special version ofsaid suspensions, said extended surface aggregates aremembrane-enclosed, liquid-filled vesicles, said first component is amembrane-forming lipid, and said second and third components aremembrane-destabilising components.

It is also an aspect of this invention to provide said suspensionswherein the third component is an NSAID, as defined above, mostpreferably is ketoprofen, ibuprofen, diclofenac, indomethacin, naproxenor piroxicam. To prepare said suspensions with these or other NSAIDingredients, the first, stable membranes forming, component is selectedfrom the group consisting of lipids, lipoids from a biological source,corresponding synthetic lipids or lipoids, or modifications thereof. Inthis context it is preferable to choose amongst glycerides, glycolipids,glycerophospholipids, isoprenoidlipids, sphingolipids, steroids,sterines or sterols, sulphur-containing lipids, lipids containing atleast one carbohydrate residue, or other polar fatty derivatives.Specifically, the preferred choice are the groups ofphosphatidylcholines, phosphatidylethanolamines, phosphatidylglycerols,phosphatidylinositols, phosphatidic acids, phosphatidylserines,sphingomyelins, sphingophospholipids, glycosphingolipids, cerebrosides,ceramidpolyhexosides, sulphatides, sphingoplasmalogenes, organgliosides.

It is a further aspect of this invention to teach that the firstsuspension component is preferably selected amongst lipids with one ortwo, not necessarily identical, fatty chains, especially with acyl-,alkanoyl, alkyl-, alkylene-, alkenoyl-, alkoxy, or chains withomega-cyclohexyl-, cyclo-propane-, iso- or anteiso-branched segments, orthe corresponding chains mixtures. Useful chains include n-decyl,n-dodecyl (lauryl), n-tetradecyl (myristyl), n-hexadecyl (palmityl),n-octadecyl (stearyl), n-eicosyl (arachinyl), n-docosyl (behenyl) orn-tetracosyl (lignoceryl), 9-cis-dodecenyl (lauroleyl),9-cis-tetradecenyl (myristoleyl), 9-cis-hexadecenyl (palmitoleinyl),9-cis-octadecenyl (petroselinyl), 6-trans-octadecenyl (petroselaidinylj,9-cis-octadecenyl (oleyl), 9-trans-octadecenyl (elaidinyl),9-cis-eicosenyl (gadoleinyl), 9-cis-docosenyl (cetoleinyl) orn-9-cis-tetracosoyl (nervonyl), n-decyloxy, n-dodecyloxy (lauryloxy),n-tetradecyloxy (myristyloxy), n-hexadecyloxy (cetyloxy), n-octadecyloxy(stearyloxy), n-eicosyloxy (arachinyloxy), n-docosoyloxy (behenyloxy) orn-tetracosoyloxy (lignoceryloxy), 9-cis-dodecenyloxy (lauroleyloxy),9-cis-tetradecenyloxy (myristoleyloxy), 9-cis-hexadecenyloxy(palmitoleinyloxy), 6-cis-octadecenyloxy, (petroselinyloxy),6-trans-octadecenyloxy (petroselaidinyloxy), 9-cis-octadecenyloxy(oleyloxy), 9-trans-octadecenyloxy (elaidinyloxy), and 9-cis-eicosenyl(gadoleinyloxy), 9-cis-docosenyl (cetoleinyloxy) or n-9-cis-tetracosoyl(nervonyloxy), n-decanoyloxy, n-dodecanoyloxy (lauroyloxy),n-tetradecanoyloxy (myristoyloxy), n-hexadecanoyloxy (palmitoyloxy) In-octadecanoyloxy (stearoyloxy), n-eicosanoyloxy (arachinoyloxy),n-n-docosoanyloxy (behenoyloxy) and n-tetracosanoyloxy (lignoceroyloxy),9-cis-dodecenyloxy (lauroleoyloxy), 9-cis-tetradecenoyloxy(myristoleoyloxy), 9-cis-hexadecenoyloxy (palmitoleinoyloxy),6-cis-octadecenoyloxy (petroselinoyloxy), 6-trans-octadecenoyloxy(petroselaidinoyloxy), 9-cis-octadecenoyloxy (oleoyloxy),9-trans-octadecenoyloxyelaidinoyloxy), and 9-cis-eicosenoyloxy(gadoleinoyloxy), 9-cis-docosenoyloxy (cetoleinoyloxy) and9-cis-tetracosenoyloxy (nervonoyloxy) or the corresponding sphingosinederivative chains, or corresponding two double bonds combinations,especially in the sequence 6,9-cis, 9,12-cis or, in case, 12,15-cis orelse the related three double bonds combinations, especially in thesequence, 6,9,12-cis, or 9,12,15-cis are preferable. A preferred choicein case of phosphatidylcholines of biological, and preferably plant,origin, is to use the lipids extracted from soy (bean), coconut, olive,safflower or sunflower, linseed, evening primrose, primrose, castor oil,and the like.

According to the invention the second suspension component, which tendsto destabilise lipid membranes, is preferably a surfactant. The selectedsurfactant can belong to the group of nonionic, zwitterionic, anionicand cationic surfactants. Preferentially, any such surfactant is chosento have solubility in the liquid medium ranging from about 5×10⁻⁷ M toabout 10⁻² M. An alternative definition of surfactants useful for theuse in said suspensions of extended surface aggregates relates tohydrophilicity-lipophilicity ratio (HLB), which should be between 10 and20, preferably between 12 and 18 and most preferred between 13 and 17. Agood choice of non-ionic surfactants according to this invention arepolyethyleneglycol-sorbitan long fatty chain esters, frompolyethyleneglycol-long fatty chain esters or -ethers and frompolyhydroxyethylen-long fatty chain esters or -ethers; preferably, thenumber of ethyleneglycol or hydroxyethylen units per such surfactantmolecule is selected to be in the range 6 to 30, more conveniently to bebetween 8 and 25 and most and typically to be between 12 to 20.Alternatively, non-ionic phospholipids with water solubility similar tothat of said non-ionic surfactants, can be used to the same effect.Examples include lysophospholipids, certain phosphatidylglycerols,phospholipids with one long and one short (C1-C6) chain, and the like.In order to ensure sufficient fluidity of resulting complex extendedsurface aggregates, the hydrophobic chain attached to such polar groupsis preferentially chosen to be sufficiently short or to be unsaturated;polyethylenglycol-sorbitan-monolaurate andpolyethylenglycol-sorbitan-monooleate, polyethyleneglycol-monolaurateand polyethyleneglycol-monooleate orpolyethyleneglycol-monolaurate-ether andpolyethyleneglycol-monooleate-ether are good choices in this respect.More specifically, it is preferable in the context of this invention, touse a surfactant which is polyethyleneglycol-sorbitan-monooleate ormonolaurate (e.g. Tween 80 or Tween 20) or else ispolyethyleneglycol-oleate or laurate (i.e. POE-oleate or POE-laurate) orelse is polyethyleneglycol-oleyl-ether or lauryl-ether, with 6 to 30,more preferably 8 to 15 and most preferred 12 to 20 ethyleneglycol (i.e.oxyethylene or OE) units per surfactant molecule.

It is another aspect of this invention to combine, in said suspension, aphosphatidylcholine as the first component and ketoprofen, diclofenac,ibuprofen, indomethacin, naproxen, or piroxicam as the third NSAIDcomponent. A preferred choice is the combination of soyphosphatidylcholine as the first and of ketoprofen, diclofenac,ibuprofen, indomethacin, naproxen or piroxicam as the third component.

In a preferred embodiment of the invention, the second component is anon-ionic surfactant, such as a polyethyleneglycol-sorbitan-long fattychain ester, a polyethyleneglycol-long fatty chain ester or apolyethyleneglycol-long fatty chain ether or else the correspondingsurfactant with a polyhydroxyethylene polar group. A preferred choice isthe use of polyethyleneglycol-sorbitan-monooleate or -laurate, ofpolyethyleneglycol-monooleate or -laurate, or else ofpolyethyleneglycol-oleyl-ether or -lauryl-ether as the second component.In the resulting suspension, the second component is preferably chosento carry a polyethyleneglycol (PEG or POE) polar head with 6 to 30, morepreferably 8 to 15 and most preferred 12 to 20 ethyleneglycol (i.e.oxyethylene or OE) units per surfactant molecule. Alternatively,non-ionic phospholipids, with water solubility similar to that of saidnon-ionic surfactants can be used for similar purpose. Moreover, thehydrophobic chains are chosen to be in a fluid state or at least to becompatible with such state of a carrier aggregate.

In another preferred embodiment of this invention is to provide saidsuspensions that contain aggregates with an average diameter before theaggregates penetrate the pores, at least 40% larger than the averagepore diameter in the barrier of interest.

In a preferred embodiment of the invention, extended surface aggregatesare proposed to have an average aggregate diameter that is at least 50%larger before pore penetration than the average pore diameter.Preferably, the average aggregate diameter before the aggregatespenetrate the pores is at least 70%, even more preferably is at least100% and most preferably is at least 150% larger than the average porediameter.

Another aspect of the invention is to provide suspensions in which thefirst component and the second component differ in solubility in theliquid medium at least 10-fold, on the average. The preferred differencein solubility between the second and third component is, on the average,at least 2-fold.

In a further preferred embodiment of the invention said suspensioncomprises a total dry mass of the at least three amphipatic componentsbetween 0.01 weight-% and 50 weight-%. A more preferred choice is tokeep this total dry mass between 0.1 weight-% and 40 weight-%, better tokeep even it between 0.5 weight-% and 30 weight-% and best to select thetotal dry mass of the three amphipatic components between 1 weight-% and15 weight-%, at the time of formulation preparation and/or application.

Yet another aspect of the invention is to provide suspensions ofextended surface aggregates, formed by the three components, with anaverage curvature corresponding to the average aggregate diameterbetween 15 nm and 5000 nm, preferably between 30 nm and 1000 nm, morepreferred between 40 nm and 300 nm and most preferred between 50 nm and150 nm.

Another aspect of the invention is to advocate using suspensions ofextended surface aggregates that contain a lower aliphatic alcohol witha membrane partition coefficient and polarity such that the alcohol, asthe at least one further second component, takes the role of a membranedestabilising component. Alcohols that potentially qualify for such useinclude mono-alcohols, diols, or to some extent polyols, of low carbonnumber (CI-C6), and ethers thereof; preferred examples are ethanol,isopropanol, 1,2-propanediol, propylene glycol, glycerol, ethyleneglycol, ethylene glycol monoethyl or monobutyl ether, propylene glycolmonomethyl, monoethyl or monobutyl ether, diethylene glycol monomethylor monoethyl ether and analogous products. The preferred choice aresimple alcohols, short chain diols or a short chain triols, preferablywith the OH-residues grouped together, corresponding methyl-, ethyl-, orbutyl-derivatives also being a possibility. This includes especiallyn-propanol, iso-propanol, or 2-propanol, n-butanol, or 2-butanol,1,2-propanediol, 1,2-butanediol; if ethanol is used, the total alcoholand lipid concentration are selected such that practically usefulethanol association with a pore penetrating aggregate is ensured.Specifically, if used individually to increase extended surfaceaggregate adaptability, ethanol, n-propanol, 2-propanol, butanol, andbenzyl alcohol are preferably used at concentrations up to 15 w-%, 10w-%, 8 w-%, 4 w-% and 2 w-%, respectively, in case of an initially 10w-% total lipid suspension. The published water-membrane partitioncoefficients for other alcohols can be used together with theserecommendations to select preferred concentration of other alcohols, orof alcohol combinations.

An important further aspect of the invention is to proposepharmaceutical preparations comprising suspensions according to theinvention. A very convenient and preferred form of aggregates in suchsuspension is that of liquid-filled vesicles in an aqueous medium, thevesicles being enclosed by membranes formed from at least one lipidcomponent and comprising at least two membrane destabilising componentsone of which is an NSAID, whereby the extended surfaces formed by thefirst and second component alone or else by the first and thirdcomponent alone, the second or third component, respectively, beingpresent at a relative concentration X, have a lower propensity toovercome barriers with pores at least 50% smaller than the averageaggregate diameter before the pore crossing than the extended surfacesformed by the first, second and third component together, if the secondand third components are present at or below the combined relativeconcentration of X.

It is also an important aspect of the invention, to teach pharmaceuticalpreparations comprising a suspension of liquid-filled vesicles in anaqueous medium, the vesicles being enclosed by membranes formed from atleast one lipid component and comprising at least three membranedestabilising components, whereby the membrane destabilising componentscomprise at least one surfactant, at least one lower aliphatic alcoholand at least one non-steroidal anti-inflammatory drug; such that themembrane destabilising components increase the adaptability of theresulting extended surface aggregates with at least three componentscompared to the aggregates containing respective combinations of thefirst and the third or the first and the second components alone.

It is a further aspect of the invention to provide pharmaceuticalpreparations comprising a suspension of liquid-filled vesicles in anaqueous medium, the vesicles being enclosed by membranes formed from atleast one lipid component and comprising at least three membranedestabilising components, whereby the membrane destabilising componentscomprise at least one surfactant, at least one lower aliphatic alcoholand at least one non-steroidal anti-inflammatory drug, such that themembrane destabilising components increase the deformability of thevesicles and the vesicles are capable of penetrating barriers with poresat least 50% smaller than the average aggregate diameter before thepenetration without changing their diameter by more than 25%.

It is another aspect of the invention to provide pharmaceuticalpreparations comprising a suspension of liquid-filled vesicles in anaqueous medium, the vesicle being enclosed by membranes formed from atleast one lipid component and comprising at least three membranedestabilising components, whereby the membrane destabilising componentscomprise a surfactant, a lower aliphatic alcohol and a non-steroidalanti-inflammatory drug, whereby the membrane destabilising componentsincrease the vesicle ability to penetrate mammalian skin and thusincrease the reach of NSAID distribution in the skin, and beyond, incomparison with the result of an NSAID application in a solution on theskin.

A preferred embodiment of the invention provides vesicle containingpharmaceutical preparations in which a phosphatidylcholine takes therole of first component and an NSAID, such as ketoprofen, diclofenac,ibuprofen indomethacin, naproxen, or piroxicam is the third component.

In another preferred embodiment of the invention pharmaceuticalpreparations contain a nonionic surfactant, preferably apolyethyleneglycol-sorbitan-long fatty chain ester, apolyethyleneglycol-long fatty chain ester or a polyethyleneglycol-longfatty chain ether, the polyethyleneglycol chain being potentiallyreplaced by a polyhydroxyethylene polar group.

Specifically preferred are polyethyleneglycol-sorbitan-monooleate (e.g.Tween 80) or -laurate (e.g. Tween 20), or elsepolyoxyethylene-monooleate (e.g. Cithrol 10MO/Chemax E-1000) or -laurate(e.g. Cithrol 10 mL) or else polyoxyethylene-oleyl-ether (e.g. Brij 98)or -lauryl-ether (e.g. Brij 35). Alternatively, non-ionic phospholipids,with water solubility similar to that of said non-ionic surfactants, areused as a preferred nonionic surfactant.

In a related embodiment of the invention, said pharmaceuticalpreparations contain an alcohol, which preferably is selected fromn-propanol, iso-propanol, 2-propanol, n-butanol or 2-butanol,1,2-propanediol, or 1,2-butanediol, a methyl- or ethyl-derivativethereof, or ethanol. When ethanol is used, the total alcohol and lipidconcentration is chosen to ensure a practically useful ethanolassociation with a pore penetrating aggregate.

It is also an aspect of the invention to provide such pharmaceuticalpreparations that are characterised by the bulk pH value above thelogarithm of the apparent dissociation constant (pKa) of the NSAID in asolution and in the extended surface aggregates, the latter pKa beinghigher than the former. Preferably, the bulk pH value is selected to bebetween 0.2 pH and 2.2 pH units above pKa of the NSAID in an extendedsurface aggregate, more preferably is between 0.5 pH and 1.9 pH unitsabove this pKa and ideally is between 0.8 pH and 1.6 pH units above suchpKa. Specifically, for the particularly interesting NSAIDs, ketoprofenor ibuprofen, the selected bulk pH is between 6.4 and 8.3, morepreferably is between 6.7 and 8 and most preferably is between 7 and7.7; for diclofenac the preferred bulk pH is between 6.2 and 8.1, morepreferably is between 6.5 and 7.8 and most preferably is between 6.8 and7.5; for naproxen the corresponding preferred pH value is between 6.3and 8.2, more preferably is between 6.6 and 7.9 and most preferably isbetween 6.9 and 7.6; for piroxicam the choice of suspension bulk pHshould be between 7.2 and 9, more preferably between 7.3 and 8.5 andmost preferably between 7.4 and 8.2.

It is another aspect of the invention to select the bulk ionic strengthof said pharmaceutical preparation is between 0.005 and 0.3, even betterbetween 0.01 and 0.2 and best between 0.05 and 0.15.

In preferred embodiment of the invention the said pharmaceuticalformulation has viscosity between 50 mPa s and 30,000 mPa s. Preferably,the formulation viscosity is chosen to be between 100 mPa s and 10,000mPa s, even better between 200 mPa s and 5000 mPa s, and most preferredbetween 400 mPa s and 2000 mPa s. To achieve such viscosity, at leastone thickening agent may be added to said pharmaceutical formulation,precise choice and concentration of such agent depending on the ambienttemperature, pH, ion strength, presence of other viscosity modifiers(such as glycerol), etc.

Thickening agents that are useful in the context of present inventionare typically pharmaceutically acceptable hydrophilic polymers,including partially etherified cellulose derivatives, such ascarboxymethyl-, hydroxyethyl-, hydroxypropyl-, hydroxypropylmethyl- ormethyl-cellulose; completely synthetic hydrophilic polymers, includingpolyacrylates, polymethacrylates, poly(hydroxyethyl)-,poly(hydroxypropyl)-, poly(hydroxypropylmethyl)methacrylate,polyacrylonitriles, methallyl-sulphonates, polyethylenes,polyoxiethylenes, polyethylene glycols, polyethylene glycol-lactides,polyethylene glycol-diacrylates, polyvinylpyrrolidones, polyvinylalcohols, poly(propylmethacrylamide), poly(propylenefumarate-eo-ethylene glycol), poloxamers, polyaspartamides, (hydrazinecross-linked) hyaluronic acids, silicone; natural gums, such asalginates, carrageenan, guar-gum, gelatine, tragacanth, (amidated)pectin, xanthan, chitosan collagen, agarose; mixtures and furtherderivatives or co-polymers thereof and/or other biologically acceptablepolymers. Most of such thickening agents in said pharmaceuticalpreparation are employed in weight concentration between 0.1 w-% and 10w-%.

For the use of pharmaceutical formulations of the invention, thefollowing hydrophilic polymer are preferred, amongst others: partiallyetherified cellulose derivatives, such as carboxymethyl-, hydroxyethyl-,hydroxypropyl-cellulose or amongst completely synthetic hydrophilicpolymer s from the class of polyacrylates, such as polymethacrylates,poly(hydroxyethyl)-, poly(hydroxypropyl)-, poly(hydroxypropylmethyl)methacrylate, especially Carbopols.

Most preferably, such formulation thickeners are chosen from the groupof polysaccharides and derivatives thereof that are commonly used on theskin, including e.g. hyaluronic acid or hydroxypropylmethylcellulose;particularly preferable choices from the group of polyacrylates includethe group of Carbopols, such as Carbopol grades 974, 980, 981, 1 382, 2984, 5 984, in each case individually or in combination. In case ofCarbopols (e.g. Carbopol 974), used to thicken the suspension-basedmulticomponent formulations for improving NSAID delivery throughpermeability barriers and the skin, the polymer concentration preferablyis selected to be between 0.3 w-% and 5 w-%, better between 0.5 w-% and3 w-% and best between 0.75 w-% and 1.75 w-%. Manufacturer'srecommendations for obtaining certain viscosity can be combined withthese guiding concentrations to use other polymers or polymercombinations in a formulation for similar purpose.

It is another preferred embodiment of the invention to use at least oneantioxidant in said pharmaceutical formulations, which is typicallyselected amongst synthetic phenolic compounds and their derivatives, thequinone-group containing substances, aromatic amines, ethylenediaminederivatives, various phenolic acids, tocopherols and their derivatives,including the corresponding amide and thiocarboxamide analogues;ascorbic acid and its salts; primaquine, quinacrine, chloroquine,hydroxychloroquine, azathioprine, phenobarbital, acetaminephen;aminosalicylic acids and derivatives; methotrexate, probucol, sulphur orphosphate atom containing anti-oxidants, thiourea; chellating agents,miscellaneous endogenous defence systems, and enzymatic antioxidants,etc. Preferred are combinations of at least two antioxidants, one beinglipophilic, such as butylated hydroxyanisol (BHA), butylatedhydroxytoluene (BHT), di-tert-butylphenol, or tertiary butylhydroquinone(TBHQ), and the other being hydrophilic, such as a chellating agent,especially EDTA, GDTA, or desferral, and/or is a sulphite, such assodium or potassium metabisulphite, a pyrosulphate, pyrophosphate orpolyphosphate. The butylated hydroxyanisol (BHA) or hydroxytoluene (BHT)are typically used at concentrations between about 0.001 w-% and about 2w-%, more preferably between about 0.0025 w-% and about 0.2 w-%, andmost preferably is between about 0.005 w-% and about 0.02 w-%; EDTA orGDTA concentration is typically chosen between about 0.001 w-% and about5 w-%, preferably between about 0.005 w-% and about 0.5 w-%, morepreferably between about 0.01 w-% and about 0.2 w-% and most preferablybetween about 0.05 and about 0.975 w-%; a sulphite, such as sodium orpotassium metabisulphite is used preferably used in concentration rangebetween about 0.001 w-% and about 5 w-%, more preferably between about0.005 w-% and about 0.5 w-%, and most preferably between about 0.01 w-%and about 0.15 w-%.

In preferred embodiments of the invention pharmaceutical preparationscontain at least one microbicide in concentration range between about0.1 w-% and about 5 w-%, as is required for proper action and as isacceptable by a regulatory body.

In presently preferred pharmaceutical preparations the first, i.e.phospholipid, component and the third, i.e. NSAID, components arepresent in the molar range between about 10/1 and about 1/1. A morepreferred range molar range of these two components is between about 5/1and about 2/1, or even between about 4/1 and about 2.5/1 and the mostpreferred composition have phospholipid/NSAID molar ratio near about3/1.

Likewise, it is preferred according to of the invention that the molarconcentration ratio of the phospholipid component, which forms stablelipid membranes, and of the second, surfactant like component, whichdestabilises such membranes, in said pharmaceutical preparations shouldbe between about 40/1 and about 4/1. More preferably such a molar ratiois between about 30/1 and about 7.5/1, the ratios between about 20/1 andabout 10/1 being most preferred.

It is a further aspect of the invention to provide a kit, comprising, ina tube or otherwise packaged form, at least one dose of thepharmaceutical preparation containing an NSAID associated with theaggregates suitable for overcoming biological barriers such as the skin.

It is another aspect of the invention to provide a method for treatingperipheral pain and/or inflammation by applying said pharmaceuticalpreparation on the skin of a warm blooded mammal.

A further aspect of the invention is to select different formulationdoses per area to control the depth of NSAID delivery, if desirableusing a non-occlusive patch for the purpose.

In a special embodiment of the invention at least one dose of an NSAIDin said pharmaceutical formulation is applied, and the application isrepeated several, e.g. up to five times per day, if necessary, thepreferred choice being two applications per day.

Last but not least, it is envisaged by the invention to use transdermalcarriers, typically in the form of barrier penetrating extended surfaceaggregates, to deliver NSAIDs below the skin and into the underlyingmuscle tissue and/or the adjacent joints.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1: Penetration curves for different SPC/KT mixtures: ·Δ·=2.5/1SPC/KT, (3/1 SPC/KT, ∇ 4/1 SPC/KT, ESPC/Tween 1/1 Transfersomes® as aReference suspension. The curves were calculated within the framework ofdata fitting model described in parallel application, by using eq. (*).

FIG. 2: Penetration curves for SPC/KT 3/1 mole/mole formulation without(o) and with (*)10 rel-mol % of Tween 80. * ReferenceTween-Transfersomes®. The curves were calculated as described in FIG. 1,using eq. (*).

FIG. 3: Area under the curve (AUC), which reflects the cumulativedelivery of the drug, calculated from the pharmacokinetic resultsmeasured with different ketoprofen (KT) formulations tested in pigs(n=4).

PRACTICAL EXAMPLES

The following examples illustrate the invention without limiting it. Alltemperatures are in degree Celsius. Carrier diameters are in nanometers,pressures are in Pascal (Pa) and other units correspond to the standardSI system. Ratios and percentages are given in moles, unless otherwisestated.

All measurements were done at room temperature, except when specifiedotherwise. For aggregate adaptability/barrier transport resistancemeasurements the test temperature was constant to within plus/minus 2degrees. For aggregate diameter measurements the temperature accuracywas plus/minus 0.1 degree. The pH value of the bulk suspension wasdetermined with a commercial (gel) electrode. Suspension viscosity wasmeasured with a rotation viscosimeter, typically at room temperature andusing 20 RPM, which corresponded to 150 1/s.

All substances were used as received and were of p.a. quality, unlessstated otherwise. Molar masses were taken to be identical to thepublished reference data.

Aggregate Adaptability Determination was conveniently conducted bymeasuring the normalised penetrability of a semi-permeable barrier totest aggregate suspension, as is described in great detail in copendingU.S. application entitled “Aggregates with increased deformability,comprising at least three amphipats, for improved transport throughsemi-permeable barriers and for the non-invasive drug application invivo, especially through the skin”, filed concurrently, the disclosureof which is already incorporated herein by above reference.

In short, aggregate adaptability is identified with the inverse pressuredifference 1/p* needed to attain a predefined, practically relevantfraction of maximum achievable suspension flux-pressure ratio (P_(max)).Using 50-60% maximum penetrability criterion gives reasonable results.Specifically, all p* values given in this document correspond to 57% ofP_(max)-value.

Exemplary results are given in FIGS. 1 and 2. The latter figure alsographically illustrates the meaning of parameters “p*” (in pressureunits, and proportional to barrier transport resistance) and “Maximumpenetrability” (=P_(max); in flux per pressure units, and indicative ofbarrier porosity).)

Aggregate diameter determination. The average aggregate (most oftenvesicle) diameter was measured with the dynamic light scattering (for afew samples with a Malvern Zeta-Sizer instrument and for the majority ofsamples with the instrument with an ALV 5000 correlator. Cumulantanalysis method are an implementation of software package “Contin” wasused for the analysis of the correlation curves obtained withZeta-Sizer. To analyse the ALV measurements the software delivered bythe manufacturer (cumulant analysis or “Contin”) was employed.

Examples 1-4 Composition

80.0-71.4 mg Phosphatidylcholine from soy-bean (SPC) 20-28.6 mgKetoprofen, sodium (KT), replacing SPC in the suspension to achieveconstant amphipat amount add 1 ml Phosphate buffer, pH = 7.2, ifnecessary readjusted with NaOH

Objective: to demonstrate that ketoprofen, an NSAID, acts as membranedestabilising component and can render mixed amphipat aggregates withextended surface adaptable enough to penetrate narrow pores.

Test suspension preparation. The stated phospholipid and drug amountswere brought into suspension using mechanical homogenisation. Thatresulting average aggregate diameter was around 100 nm. For reference, acomparable suspension containing SPC and sodium cholate in 3.75/1mol/mol ratio was used.

Vesicle transport ability (pore penetration capability/adaptability).The efflux of the test suspension from a vessel pressurised withnitrogen gas was measured as a function of the time to determine thepressure dependency of material transport through the 20 nm pore filterin front of an opening in the measuring vessel. From the measured fluxdata, the effective “barrier penetrability”, which defines theadaptability of the tested mixed amphipat vesicles, was calculated as isdescribed in the main text body. The measured curves were also analysedin terms of the pressure p*, needed to achieve 57% of maximum possiblesuspension flux/pressure ratio. The calculated p*-value decreased from2.41±0.15 MPa (mean value k standard error) through 1.66 k 0.07 MPa to1.36±0.10 MPa with increasing drug concentration. This is indicative ofmembrane destabilising activity of the drug, which arguably promotesbilayer flexibility and permeability. More detailed information is givenin Table 1, which reveals essentially identical p* values for the SPC/KT3/1 mol/mol mixture and for the reference anionic Transfersome®suspension. To deduce vesicle adaptability from p*-value, contributionfrom suspension viscosity effects must be included or must be known tobe negligible. This is not an issue, however, as long as one can makecomparisons with suitable reference formulation(s), as is done in thefollowing table by inclusion of the last row.

In this test series, and in all other practical examples reportedherein, the final aggregate diameter after narrow pore crossing was atleast 300%, and typically was more than 400% greater than the porediameter, the final to starting aggregate diameter ratio beingtypically >0.70, implying fragmentation of less than 30%, and more oftenmerely 10-20%.

TABLE 1 Fit results, based on eq. (*) for the barrier penetrability(flux/pressure ratio) experiments one with the suspensions characterisedby different lipid/drug, SPC/KT ratios SPC/KT p* P_(max) Adaptability[mole/mole] [MPa] [10⁻¹¹ mPA⁻¹ · sec⁻¹] a_(a), [MPa⁻¹] 10/0  ~3 Notmeasurable ~0.3 4/1 2.41 ± 0.15^(§) Not measurable 0.145 3/1 1.66 ±0.07^(§) — 0.602 2.5/1   1.36 ± 0.10^(§) 345 ± 37 0.735 Referenceanionic 1.76 ± 0.13^(§) 318 ± 39 0.568 Tfs^(§§) ^(§)The quoted erroronly accounts for analytical and not for experimental data uncertainty,the latter often amounting to 20-30%. ^(§§)These Tfs vesicles wereprepared from an SPC/Na cholate 3.75/1 mol/mol mixture.

Graphic representation of the results of these experiments is given inFIG. 1.

Examples 5-7 Composition

75.0, 75.0, 37.7 mg Phosphatidylcholine from soy-bean (SPC) 25.0, 25.0,0.0 mg Ketoprofen, sodium (KT) 0.0, 25.4, 62.3 mg Tween 80 0.0, 0.0,37.7 mg Ethanol add 1 ml Phosphate buffer (pH = 7.2)

Objective: to test the synergistic effect of the second and firstmembrane destabilising amphipat (Tween 80, ketoprofen, respectively) interms of an extended surface aggregate adaptability.

Suspension preparation was essentially the same as with examples 1-4.

Vesicle transport ability (pore penetration capability/adaptability).Transbarrier flux of the test suspension containing 5 mol-% Tween ismuch higher than for the formulation that contains merely phospholipid(as the basic amphipat) and ketoprofen (as the surface active, membranedestabilising, surfactant-like amphipat) components. This is clearlyseen from FIG. 2, which illustrates pressure dependence of saidsuspension flux divided by driving pressure.

Examples 8-12 Composition of Aggregates

75.0 mg Phosphatidylcholine from soy-bean (SPC), the actual value is: 75mg-Tween 80 amount in mg 25.0 mg Ketoprofen, sodium (KT) see the Tween80 following table add 1 ml Phosphate buffer (pH = 7.2)

Reference buffer: Phosphate buffer (pH=7.2)

Objective: to study the effect of relative concentration of asurfactant, as the second membrane destabilising amphipat, onadaptability of extended surface mixed amphipat aggregates.

Suspension preparation: as with examples 1-4.

Vesicle transport ability (pore penetration capability/adaptability)data, as measured in this test series, confirm and expand the findingsobtained with examples 1-4. Tween acting as the second membranedestabilising component improves the ability of test suspension topenetrate barriers even when this surfactant is present in thequaternary mixture merely in small amount, as long as relativeconcentration of Tween is at least approximately 2.5 mol-%, and evenbetter 5 mol-%. Data given in Table 2 justify the conclusion. They arecompared with the reference non-ionic Tween-based Transfersome®formulation (Reference Tfs) and with the buffer fluid (Reference fluid)in which mixed amphipat vesicles were suspended.

The suspension viscosity for example 11 was around 730 mPa s at 20 RPM

TABLE 2 Fit results for the pore penetration experiments done withvarious quaternary suspensions of a phospholipid (SPC; stable membranesforming component), a drug (KT; 1^(st) membrane destabilisingcomponent), and Tween 80 (2^(nd) membrane destabilising component)co-suspended in a buffer at different relative concentrations of Tween80. P_(max) Tween 80 content p* [10⁻¹¹ Adaptability Nr [mol % of SPC][MPa] mPA⁻¹ sec⁻¹] a_(a), [MPa⁻¹] 0 1.66 ± 0.07 345 ± 37 0.602 8 1.250.51 ± 0.05 293 ± 23 1.961 9 2.5 0.50 ± 0.04 339 ± 26 2.000 10 5 0.23 ±0.03 215 ± 19 4.348 11 7.5 0.22 ± 0.02 213 ± 14 4.545 12 Reference Tfs0.20 ± 0.01 227 ± 3  5.000 (Tween) Reference fluid Not applicable 613 ±15 Not applicable (buffer) ^(§)The quoted error only accounts foranalytical and not for experimental data uncertainty, the latter oftenamounting to 20-30%. Reference Tfs vesicles were prepared from anequimolar (50/50 mol/mol) SPC/Tween 80 mixture.

Comparative Examples 13-14 Composition

43.65 mg Phosphatidylcholine from soybean (+95% = PC) 72.00 mg Tween 8034.35 mg Ketoprofen 6.08 mg Sodium hydroxide 5.25 mg Benzyl alcohol36.51 mg Ethanol 96% add 1 g 154 mM phosphate buffer, pH = 7.4

Suspension preparation: Formulations were made in accordance with theabove compositions.

Hydration of the components mixed in given proportions produced a clear,yellowish fluid. This is indicative of micellar suspension and impliesthat the tested mixed lipid aggregates are colloidally not stable.Determination of the average diameter of aggregates in such suspensionwith the dynamic light scattering confirmed the conclusion (meanparticle diameter approx. 22 nm, which is incompatible with existence ofvesicles).

Diluting the preparation with the corresponding buffer from 15% totallipid to 10% total lipid, making essentially the same observationfurther corroborated the result. Based on the existing information aboutphosphatidylcholine solubilisation by Tween 80, even a reduction ofrelative surfactant concentration by a factor of 2, thus creating aSPC/Tween 80 2/1 mol/mol mixture loaded with approx. 30 mol-%ketoprofen, still would yield unstable aggregates.

Addition of Tween 80 much beyond the rather low relative molarconcentration proposed in example 12 thus destabilises the threecomponent lipid aggregates to the point of solubilisation, or at closeto this point. Such compositions, therefore, do not fulfil the requiredstability criterion for the extended surface aggregates required by thepresent application.

Comparative Examples 15-16 Composition

66.71 mg Soybean-phosphatidylcholine 11.00 mg Tween 80 22.21 mgKetoprofen 0.00/66.71 mg Ethanol (EtOH; for examples 16 and 17,respectively) 11.56 mg NaOH (30%) 0.50 mg Na metabisulphite 1.00 mgDisodium edetate (EDTA) 0.20 mg Butylhydroxytoluene (BHT) 1.46 mgMethylparabene 1.00 mg Linalool 5.25 mg Benzyl alcohol add 1 g 7.8 mMPhosphate buffer, pH = 7.2

Suspension preparation. Vesicular intermediate preparation with 17.14%total lipid containing no ethanol and ketoprofen in identicalconcentration as in Example 11 was mixed with the SPC mass equivalent ofethanol. To meet the needs of pharmaceutical formulations as well,several suspension stabilising agents (EDTA, BHA, methylparabene, andbenzyl alcohol) were included in the formulation. Characterisation wasdone as with examples 1-4.

TABLE 3 Results of driving pressure and aggregate adaptability analysisfor examples 15 and 16. p* Pmax Adaptability Formulation [MPa] [10⁻⁸kg/(m² · s · Pa)] aa₁ [MPa⁻¹] Example 15 (no 0.233 ± 0.013^(§) 216.5 ±7.4 4.292 EtOH) Example 16 (with 0.133 ± 0.006^(§) 254.3 ± 91  7.519EtOH) ^(§)The quoted error only accounts for analytical and not forexperimental data uncertainty.

Specifically, the pressure required to drive vesicles through narrowpores, p*, was found to decrease in the presence of EtOH from 0.233 MPato 0.133 mPa; this is a decrease of approx. 40% and thus near the limitof insignificance (see Table 2 for comparison). The reason is thelimited assay resolution, which for p* in the studied situation is20-30%.

Speaking in absolute terms, and making comparison with the magnitude ofpositive effect on aggregate adaptability caused by Tween 80 (cf. Tables2 and 3), ethanol in the ranges tested only increases the adaptabilityof tested aggregates moderately.

Comparison of the results from experiments 15 and 16 and 11, moreover,confirms that the tested system preservatives (Na metabisulphite; EDTA;BHT, benzyl alcohol) neither affect negatively the desirable extendedsurface aggregate adaptability nor do they change much the pressurerequired for driving adequate suspension transport through a nano-porousbarrier.

Further, the results, given in Table 3, confirm that the adaptability ofthe aggregates proposed in the Comparative Examples is far inferior tothat of the present formulations.

Examples 19-21 Composition

75 mg Phosphatidylcholine from soy-bean (SPC), 25 mg Ketoprofen, sodium(KT) See the Tween 80 (mol-% referring to SPC) following table add 1 mlWater or 50 mM buffer (pH = 7.2)

Objective: to test the influence of ionic strength of the bulk inorganicelectrolyte on the adaptability of mixed amphipat aggregates suspendedin such an electrolyte.

Suspension preparation and characterisation. The test suspension wasprepared essentially as with examples 1-4, except in that the buffer wassometimes exchanged for water with practically the same pH-value. Thishad important consequences. When the ionic strength (I) of the bulkelectrolyte solution with a pH near 7 changes, ketoprofen distributionand degree of ionisation in Transfersome® suspension also changes. Thismodifies—most probably decreases—extended surface aggregateadaptability, which must be considered when designing products on thebasis of given formulation composition. Experimental evidence for thisis given in Table 5.

TABLE 5 The fit results based on formula (*) for the transbarrierfluxldriving pressure ratio (barrier penetrability), of variousquinternary suspensions with KT as the drug in different buffer systems.Pmax [10⁻¹¹ p* mPa⁻¹ · Adaptability Formulation [MPa] sec⁻¹] aa₁ [MPa⁻¹]10 mol-% Tween 0.49 ± 0.02 212 ± 8  2.041 no buffer 10 mol-% Tween, 0.25± 0.03 230 ± 17 4.000 50 mM buffer, I = 117 mM 7.5 mol-% Tween, 6.3% v/v0.31 ± 0.06 194 ± 23 3.226 EtOH no buffer 7.5 mol-% Tween, 6.3% v/v 0.13± 0.01 248 ± 11 7.692 EtOH 50 mM buffer, I = 117 mM Reference Tween Tfsin the 0.20 ± 0.01 227 ± 3  5.000 buffer

Examples 22-23 Composition

75.0 mg Phosphatidylcholine from soy-bean (SPC), 25 mg Ketoprofen,sodium (KT) 12.4 mg Tween 80 add 1 ml Buffer pH = 7.2 and pH = 7.7

Suspension preparation and characterisation: see previous test series.

Objective: to test the effect of ketoprofen ionisation, which above thepKa(KT)˜6.4 increases with pH, on adaptability of the drug loaded mixedlipid vesicles.

Results: Adaptability of simple formulations containing three amphipaticcomponents was confirmed to depend on the ionisation state of its onlytitratable component, ketoprofen. Detailed results are given in thefollowing Table 6.

TABLE 6 Fit results, based on eq. (*), for the pressure normalisedtransbarrier flux of KT-Tfs suspensions at different pH p* PmaxAdaptability pH [MPa] [10⁻¹¹ mPa⁻¹ · sec⁻¹] a_(a) [MPa⁻¹] 7.2 1.66 ±0.07 345 ± 37 0.602 7.7 0.62 ± 0.07 237 ± 28 1.613 Reference Tfs 0.20 ±0.01  227 ± 2.9 5.000

Examples 24-25 Composition

100 mg/ml Phosphatidylcholine from soy-bean (SPC) as large unilamellarvesicle suspension 254 mg/ml Ketoprofen, sodium (KT) in solution BufferpH = 7.2 and pH = 7.7 Mixed during experiments to yield increasingrelative ratio of KT in SPC aggregates suspension.

Objective: to test the ability of ketoprofen to solubilise lipid bilayermembranes.

Results: The ability of ketoprofen to solubilise soybeanphosphatidylcholine (SPC) membranes was determined by measuring theturbidity of a suspension (10 w-%) of large unilamellar vesicles duringsuccessive addition of 1 M solution of ketoprofen. In the first testseries this was done in 50 mM phosphate buffer at pH=7.4, where morethan 50% of the drug is ionised and more than 50% of the drug isvesicle-bound, but chiefly in the non-charged form, which does notdestabilise lipid membranes significantly. SPC vesicles under testedthese conditions were not measurably solubilised, despite the presenceof some ionised ketoprofen, but were partly destabilised, asdemonstrated in previous examples.

The second experiment was performed at pH=11.6, under which conditionall ketoprofen molecules are deprotonated and hence have a maximumsolubilisation, i.e. membrane destabilisation, capability.Solubilisation of SPC membranes was now observed when the molar ratiofor the drug in vesicle bilayers was above ketoprofen/SPC 10.8/1mole/mole. SPC-ketoprofen association thus produces weakly boundcomplexes with membrane solubilising capability.

Examples 26-30 Composition

75.0 mg Phosphatidylcholine from soy-bean (SPC, used as a saturatedethanolic solution) the actual number is: 75 mg - Brij content 25.0 mgKetoprofen, sodium (KT) See the following table Brij 98 add 1 mlPhosphate buffer (pH = 7.2)

Objective: to demonstrate the usefulness of another surfactant, Brij,different from Tween 80, to increase the flux through narrow pores ofketoprofen/SPC extended surface aggregates in a suspension.

Suspension preparation was essentially the same as in examples 1-4.

Flux determination. The flux of suspension of extended surfaceaggregates containing SPC, KT and, in case, Brij 98 was measured usingthe same device as is used for aggregate adaptability determination. Theonly difference was that only a single driving pressure was used forsuspension characterisation. For comparison, the ratio of KT-loaded andof empty Brij Transfersomes® was calculated (=Rel. Flux).

The results of the test series measured with Brij 98, apolyoxyethylene-oleyl-ether with 20 OE units in polar head are given inTable 7.

TABLE 7 Flux of mixed amphipat suspensions through 20 nm pores in asemi- permeable barrier driven by trans-barrier pressure of 0.1 Mpa.Brij 98 content Flux [mol % of SPC] [mg cm⁻²sec⁻¹] Rel. Flux 0 <1 2.510 >10 5.0 30 >30 7.5 29 >29

Examples 31-34 Composition KT Form(ulation) B (Expt 31)

Weight-% 2.857 Ketoprofen (USP) 7.143 Phosphatidylcholine 3.000 Glycerol(USP) 2.087 Sodium Hydroxide, 50% (FCC) 0.120 Phosphate buffer salts0.100 Linalool 0.100 Disodium edetate EDTA 1.250 Carbomer 974 0.100Carbomer 1342 1.000 Propylen Glycol 0.200 Ethylparaben 0.525 BenzylAlcohol 0.020 Butylated hydroxytoluene 81.499 Water

Composition KT Form.(ulation) A (Expt 32)

Weight-% 2.290 Ketoprofen 6.870 Soy Phosphatidylcholine (SPC) 0.850Polysorbate (Tween 80) 3.651 Ethanol 96% 0.930 NaOH (sodium hydroxide)0.235 Phosphate buffer salts 0.050 Sodium metabisulphite 0.020Butylhydroxytoluene (BHT) 0.100 Disodium edetate (EDTA) 0.250 Methylparahydroxybenzoate 0.525 Benzyl alcohol 0.100 Linalool 1.250 Carbomer(Carbopol 980) 3.00 Glycerol 79.879 Water

Commercial topical formulation Gabrilen (Expt. 33): according to deskphysicians' reference, the preparation contains 25 mg KT/g gel,supplemented with 96% ethanol, 3-propanol, 10% ammonia solution andCarbomer in purified water.

Commercial oral formulation Ketoprofen Ratiopharm (KT Ratiopharm) (Expt.34): according to desk physicians' reference each film tablet contains50 mg KT in addition to microcrystalline cellulose, gelatine, SiO₂, cornstarch, talcum, crosscarmelose sodium, Mg stearate, hypromelose,macrogol, glycerol, dyes E 171 and E 172.

Preparation of formulations A and B, which both contained extendedsurface vesicles, was done essentially as described for examples 1-4.Commercial comparators were purchased in a local pharmacy and used asobtained.

Methodology: The test pigs were numbered and central vein catheters wereimplanted into the animals. The application area on a hind limb of eachanimal was shaved with an electric clipper and cleaned with warm waterand soap. Then, an application area of 10 cm×10 cm (=100 cm²) wasmarked.

At time zero of the sampling period, 2 ml of the blood were sampled fromeach test animal into a citrate-coated vial to generate plasma. The pigswere anaesthetised for approximately 60 min and the appropriate dose ofthe test medication was applied onto the application site of a pig orelse was given to the animal orally. Further plasma samples (0.5 mleach) were taken 0.5, 1, 2, 3, 5, 8 and 12 hours post application. Theywere kept frozen until analysis.

Ketoprofen concentration was determined with HPLC using standardmethods, in case of muscle tissue samples after the specimenhomogenisation. Area under the curve (AUC) was calculated by integratingall time-point data.

Results of experiments are given in Tables 8 and FIG. 3. Whereas theindividual pharmacokinetic data sets are rather scattery, yieldingstandard deviations comparable to the mean because of small group size,the overall data analysis does demonstrate the superiority of at leastthree amphipat component preparations, in comparison with two amphipatcomponent formulations, to deliver an NSAID (ketoprofen) deep under theapplication site on the skin. The greater is the investigated tissuedepth the greater is the observed advantage (superficial muscle=0-1.5cm; deep muscle >1.5 cm).

TABLE 8a Area under the curve (AUC_(0-8 h) [ng × mg⁻¹ × h]), measuredwith different KT formulations in pigs Formulation KT Gabrilen ® BFormulation A Ratiopharm ® (n = 4) (n = 7) (n = 7) (oral, n = 3)Superficial 102 209 306 7 muscle tissue Deep 53 147 301 9 muscle tissue

TABLE 8b Ketoprofen (KT) concentration in superficial muscle tissue(ng/mg) KT-Tfs KT-Ratiopharm Time Gabrilen ® KT-Tfs Form. B Form. A(oral) (hours) (n = 4) (n = 7) (n = 7) (n = 3) 0 0.0 ± 0.0 0.0 ± 0.0 0.0± 0.1 0.0 ± 0.0 1 5.0 ± 3.3 50.4 ± 48.6 55.5 ± 66.3 1.0 ± 1.2 2 12.8 ±22.6 75.2 ± 83.8 36.3 ± 32.1 1.6 ± 1.2 3 10.9 ± 11.5 3.0 ± 3.2 25.7 ±28.5 1.4 ± 0.3 5 19.3 ± 18.7 12.9 ± 11.1 45.2 ± 72.9 0.7 ± 0.2 8 3.8 ±3.8 19.6 ± 17.9 22.0 ± 17.9 0.2 ± 0.1

TABLE 8c Ketoprofen (KT) concentration in deep muscle tissue (ng/mg)KT-Tfs KT-Ratiopharm Time Gabrilen ® KT-Tfs Form. B Form. A (oral)(hours) (n = 4) (n = 7) (n = 7) (n = 3) 0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.10.0 ± 0.0 1 2.6 ± 2.3 53.4 ± 66.5 24.8 ± 19.0 1.5 ± 1.6 2 5.4 ± 9.3 63.0± 51.9 18.8 ± 21.5 1.8 ± 1.0 3 9.0 ± 9.3 1.4 ± 0.8 49.8 ± 71.8 1.6 ± 0.55 7.9 ± 5.8 5.6 ± 2.2 49.9 ± 65.0 1.0 ± 0.2 8 2.9 ± 2.9 14.1 ± 10.9 30.2± 28.7 0.3 ± 0.2

Examples 35-36 Composition for Ketoprofen in Carrier Suspension (KT-TfsSol)

Weight-% 3.435 Ketoprofen (KT) 10.305 Soy Phosphatidylcholine (SPC)1.275 Polysorbate (Tween 80) 5.477 Ethanol 96% 0.533 NaOH (sodiumhydroxide) 0.235 Phosphate buffer salts 0.050 Sodium metabisulphite0.020 Butylhydroxytoluene (BHT) 0.100 Disodium edetate (EDTA) 0.250Methyl parahydroxybenzoate 0.525 Benzyl alcohol 0.100 Linalool 3.00Glycerol 74.695 Water

Composition for Ketoprofen in Carrier Gel (KT-Tfs Gel)

As in experiment 35, except in that the first four components arediluted 1.5-fold and Carbomer (Carbopol 980), buffered to pH=7.2, isincluded to final concentration of 1.25 w-%.

Objective: to test the effect of formulation viscosity, and the presenceof a thickening agent as viscosity modifier, on the ability of NSAIDloaded extended surface aggregates to deliver the drug (ketoprofen) deepunder the application site on the skin.

Methodology was the same as in experiments 31-34, except in that no oralcomparator was included. A total of 4 pigs were used in each group.

Area under the curve (AUC) was calculated by integrating all PK(pharmacokinetic) data measured in different tissues (plasma, not shown)and the muscles under drug application site on the skin. The resultsobtained for superficial (0-1.5 cm) and deep (>1.5 cm) muscle are givenin Tables 9, and suggest no detrimental effect of the thickening agentsused in KT-Tfs gel to achieve the desired suspension viscosity ofapprox. 730 mPa s. If anything, the thickening agent present in thetested gel is beneficial.

TABLE 9a Area under the curve (AUC_(0-8 h) [ng × mg⁻¹ × h]), measuredwith two carrier-based ketoprofen (KT) formulations in pigs KT-TfsKT-Tfs gel sol. KT-Tfs gel KT-Tfs sol. 17 mg 17 mg 50 mg 50 mg (n = 4)(n = 4) (n = 4) (n = 4) Superficial muscle 147 44 278 186 tissue Deepmuscle tissue 97 63 266 202

TABLE 9b KT concentration in superficial muscle tissue (ng/mg) KT-Tfsgel KT-Tfs sol. KT-Tfs gel KT-Tfs sol. Time 17 mg 17 mg 50 mg 50 mg(hours) (n = 4) (n = 4) (n = 4) (n = 4) 0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.10.0 ± 0.0 1 83.3 ± 82.9 2.3 ± 1.5 55.5 ± 66.3 23.0 ± 29.3 2 24.1 ± 27.50.8 ± 0.3 36.3 ± 32.1 21.2 ± 33.6 3 8.1 ± 8.0 2.8 ± 0.1 25.7 ± 28.5 9.0± 2.1 5 14.2 ± 14.2 10.6 ± 12.5 45.2 ± 72.9 34.8 ± 49.8 8 3.1 ± 2.6 3.5± 2.4 22.0 ± 17.9 29.8 ± 50.1

TABLE 9c KT concentration in deep muscle tissue (ng/mg) KT-Tfs gelKT-Tfs sol. KT-Tfs gel KT-Tfs sol. Time 17 mg 17 mg 50 mg 50 mg (hours)(n = 4) (n = 4) (n = 4) (n = 4) 0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.1 0.0 ±0.0 1 36.0 ± 49.1 14.7 ± 1.5  24.8 ± 19.0 24.5 ± 44.7 2 19.4 ± 23.5 0.8± 0.3 18.8 ± 21.5 4.5 ± 4.0 3 2.4 ± 2.6 9.2 ± 3.1 49.8 ± 71.8 25.4 ±43.0 5 13.5 ± 8.8   9.3 ± 12.5 49.9 ± 65.0 46.6 ± 85.6 8 2.4 ± 1.4 6.4 ±2.4 30.2 ± 28.7 15.6 ± 23.4

1-37. (canceled)
 38. A method for inducing analgesia comprising applyingto the skin of a warm blooded mammal a vesicular compositioncomprising: 1) vesicles having a lipid bilayer and comprising: i) aphosphatidylcholine; ii) a polyethyleneglycol-sorbitan-10 to 24 carbonatom fatty chain ester, a polyethyleneglycol-10 to 24 carbon atom fattychain ether, or a nonaethyleneglycol octylphenyl ether surfactant; andiii) an NSAID that has an acid group that is in its salt form; and 2) apharmaceutically acceptable, polar liquid medium, wherein thephosphatidylcholine and the surfactant of the vesicles are present in amolar ratio of between about 40/1 and about 7.5/1, the lipid bilayer isin the fluid lamellar phase, and the pH of the composition is above thepKa of the NSAID.
 39. The method of claim 38, wherein the NSAID isketoprofen, ibuprofen, diclofenac, indomethacin, or naproxen.
 40. Themethod of claim 38, wherein the phosphatidylcholine is from soy bean,coconut, olive, safflower, or sunflower, linseed, evening primrose,primrose, or castor oil.
 41. The method of claim 38, wherein the totaldry mass of the phosphatidylcholine, the surfactant, and the NSAID isbetween 0.01 weight-% and 50 weight-% of the composition.
 42. The methodof claim 38, wherein the composition further comprises a lower aliphaticalcohol.
 43. The method of claim 42, wherein the alcohol is n-propanol,isopropanol, 2-propanol, n-butanol, 2-butanol, 1,2-propanediol,1,2-butanediol, or ethanol.
 44. The method of claim 38, wherein the pHof the composition is between 6.4 and 8.3.
 45. The method of claim 38,wherein the ionic strength of the composition is between 0.005 and 0.3.46. The method of claim 38, wherein the viscosity of the composition isbetween 50 mPa s and 30,000 mPa s.
 47. The method of claim 38, whereinthe composition is applied in a non-occlusive patch.
 48. The method ofclaim 38, wherein the NSAID is ketoprofen, ibuprofen, diclofenac,naproxen, indomethacin, acetaminophen, diflunisal, etodolac,flurbiprofen, mefenamic acid, salsalate, tiaprofenic acid; or the saltform is choline salicylate-Mg salicylate, fenoprofen calcium, ketorolactromethamine, magnesium salicylate, sodium salicylate, or tolmetinsodium.
 49. The method of claim 38, wherein the total dry mass of thepolyethyleneglycol-sorbitan-10 to 24 carbon atom fatty chain ester,polyethyleneglycol-10 to 24 carbon atom fatty chain ether, ornonaethyleneglycol octylphenyl ether-surfactant; and the NSAID isbetween 0.01 weight-% and 50 weight-%.
 50. The method of claim 38,wherein the phosphatidylcholine is soy phosphatidylcholine or egglecithin.
 51. The method of claim 38, wherein the NSAID is ketoprofen.52. The method of claim 38, wherein the surfactant is apolyethyleneglycol-sorbitan-10 to 24 carbon atom fatty chain ester. 53.The method of claim 52, wherein the polyethyleneglycol-sorbitan-10 to 24carbon atom fatty chain ester is polyethyleneglycol-sorbitan-monooleate.54. The method of claim 53, wherein the polyethyleneglycol-sorbitan-10to 24 carbon atom fatty chain-ester is Tween
 80. 55. The method of claim38, wherein the phosphatidylcholine is soy phosphatidylcholine, thesurfactant is polyethyleneglycol-sorbitan-monooleate, and the NSAID isketoprofen.
 56. The method of claim 38, wherein the molar ratio ofphosphatidylcholine to NSAID is between about 3/1 to about 1/1.
 57. Themethod of claim 56, wherein the molar ratio of phosphatidylcholine toNSAID is between about 2.5/1 to about 1/1.
 58. The method of claim 57,wherein the molar ratio of phosphatidylcholine to NSAID is between about2/1 to about 1/1.
 59. The method of claim 58, wherein the molar ratio ofphosphatidylcholine to NSAID is about 1/1.
 60. The method of claim 55,wherein the molar ratio of phosphatidylcholine to NSAID is between about3/1 to about 1/1.
 61. The method of claim 60, wherein the molar ratio ofphosphatidylcholine to NSAID is between about 2.5/1 to about 1/1. 62.The method of claim 61, wherein the molar ratio of phosphatidylcholineto NSAID is between about 2/1 to about 1/1.
 63. The method of claim 62,wherein the molar ratio of phosphatidylcholine to NSAID is about 1/1.64. The method of claim 52, wherein the polyethyleneglycol-sorbitan-10to 24 carbon atom fatty chain ester ispolyethyleneglycol-sorbitan-monolaurate.
 65. The method of claim 64,wherein the polyethyleneglycol-sorbitan-monolaurate is Tween
 20. 66. Themethod of claim 38, wherein the pH of the composition is between 0.2 and2.2 pH units above the pKa of the NSAID.
 67. The method of claim 66,wherein the pH of the composition is between 0.5 and 1.9 pH units abovethe pKa of the NSAID.
 68. The method of claim 67, wherein the pH of thecomposition is between 0.8 and 1.6 pH units above the pKa of the NSAID.69. The method of claim 38, wherein the phosphatidylcholine is from eggor soya beans.
 70. The method of claim 38, wherein the surfactant is apolyethyleneglycol-sorbitan-monolaurate, apolyethyleneglycol-sorbitan-monooleate, apolyethyleneglycol-monolaurate, a polyethyleneglycol-monooleate, apolyethyleneglycol-monolaurate-ether, or apolyethyleneglycol-monooleate-ether.
 71. The method of claim 38, whereinthe composition further comprises a thickening agent, an antioxidant, ora microbicide.
 72. The method of claim 38, wherein thephosphatidylcholine and the surfactant are present in a molar ratio ofbetween about 40/1 and about 10/1.
 73. The method of claim 38, whereinthe phosphatidylcholine and the surfactant are present in a molar ratioof between about 30/1 and about 7.5/1.
 74. The method of claim 73,wherein the phosphatidylcholine and the surfactant are present in amolar ratio of between about 20/1 and about 10/1.
 75. The method ofclaim 74, wherein the pH is between 0.2 and 2.2 pH units above the pKaof the NSAID.
 76. The method of claim 38, wherein the NSAID isibuprofen.
 77. The method of claim 38, wherein the NSAID is diclofenac.78. The method of claim 38, wherein the NSAID is indomethacin.
 79. Amethod for inducing analgesia comprising applying to the skin of a warmblooded mammal a vesicular composition comprising: 1) vesiclescomprising: i) a phosphatidylcholine; ii) apolyethyleneglycol-sorbitan-10 to 24 carbon atom-fatty chain ester, apolyethyleneglycol-10 to 24 carbon atom-fatty chain ether, or anonaethyleneglycol octylphenyl ether surfactant; and iii) an NSAID thathas an acid group that is in its salt form; and 2) a pharmaceuticallyacceptable, polar liquid medium, wherein the phosphatidylcholine and thesurfactant of the vesicles are present in a molar ratio of between about40/1 and about 7.5/1, the vesicles are capable of penetrating a barrierwith pores having an average pore diameter at least 50% smaller than theaverage vesicle diameter before the penetration, and the pH of thecomposition is above the pKa of the NSAID.
 80. The method of claim 79,wherein the phosphatidylcholine is from soy bean, coconut, olive,safflower, or sunflower, linseed, evening primrose, primrose, or castoroil.
 81. The method of claim 79, wherein the total dry mass of thephosphatidylcholine, the surfactant, and the NSAID is between 0.01weight-% and 50 weight-% of the composition.
 82. The method of claim 79,wherein the composition further comprises a lower aliphatic alcohol. 83.The method of claim 82, wherein the alcohol is n-propanol, isopropanol,2-propanol, n-butanol, 2-butanol, 1,2-propanediol, 1,2-butanediol, orethanol.
 84. The method of claim 79, wherein the pH of the compositionis between 6.4 and 8.3.
 85. The method of claim 79, wherein the ionicstrength of the composition is between 0.005 and 0.3.
 86. The method ofclaim 79, wherein the viscosity of the composition is between 50 mPa sand 30,000 mPa s.
 87. The method of claim 79, wherein the NSAID isketoprofen, ibuprofen, diclofenac, naproxen, indomethacin,acetaminophen, diflunisal, etodolac, flurbiprofen, mefenamic acid,salsalate, tiaprofenic acid; or the salt form is choline salicylate-Mgsalicylate, fenoprofen calcium, ketorolac tromethamine, magnesiumsalicylate, sodium salicylate, or tolmetin sodium.
 88. The method ofclaim 79, wherein the NSAID is ketoprofen, ibuprofen, diclofenac,naproxen, or indomethacin.
 89. The method of claim 79, wherein the totaldry mass of the phosphatidylcholine; the polyethyleneglycol-sorbitan-10to 24 carbon atom-fatty chain ester, polyethyleneglycol-10 to 24 carbonatom-fatty chain ether, or nonaethyleneglycol octylphenyl ethersurfactant; and the NSAID is between 0.01 weight-% and 50 weight-%. 90.The method of claim 79, wherein the phosphatidylcholine is soyphosphatidylcholine or egg lecithin.
 91. The method of claim 79, whereinthe NSAID is ketoprofen.
 92. The method of claim 79, wherein thesurfactant is a polyethyleneglycol-sorbitan-10 to 24 carbon atom-fattychain ester.
 93. The method of claim 92, wherein thepolyethyleneglycol-sorbitan-10 to 24 carbon atom-fatty chain ester ispolyethyleneglycol-sorbitan-monooleate.
 94. The method of claim 93wherein the polyethyleneglycol-sorbitan-10 to 24 carbon atom-fattychain-ester is Tween
 80. 95. The method of claim 92, wherein thepolyethyleneglycol-sorbitan-10 to 24 carbon atom-fatty chain ester ispolyethyleneglycol-sorbitan-monolaurate.
 96. The method of claim 95,wherein the polyethyleneglycol-sorbitan-monolaurate is Tween
 20. 97. Themethod of claim 79, wherein the molar ratio of phosphatidylcholine toNSAID is between about 3/1 to about 1/1.
 98. The method of claim 97,wherein the molar ratio of phosphatidylcholine to NSAID is between about2.5/1 to about 1/1.
 99. The method of claim 98, wherein the molar ratioof phosphatidylcholine to NSAID is between about 2/1 to about 1/1. 100.The method of claim 99, wherein the molar ratio of phosphatidylcholineto NSAID is about 1/1.
 101. The method of claim 79, wherein thephosphatidylcholine is soy phosphatidylcholine, the surfactant ispolyethyleneglycol-sorbitan-monooleate, and the NSAID is ketoprofen.102. The method of claim 101, wherein the molar ratio ofphosphatidylcholine to NSAID is between about 3/1 to about 1/1.
 103. Themethod of claim 102, wherein the molar ratio of phosphatidylcholine toNSAID is between about 2.5/1 to about 1/1.
 104. The method of claim 103,wherein the molar ratio of phosphatidylcholine to NSAID is between about2/1 to about 1/1.
 105. The method of claim 104, wherein the molar ratioof phosphatidylcholine to NSAID is about 1/1.
 106. The method of claim79, wherein the pH of the composition is between 0.2 and 2.2 pH unitsabove the pKa of the NSAID.
 107. The method of claim 106, wherein the pHof the composition is between 0.5 and 1.9 pH units above the pKa of theNSAID.
 108. The method of claim 107, wherein the pH of the compositionis between 0.8 and 1.6 pH units above the pKa of the NSAID.
 109. Themethod of claim 79, wherein the phosphatidylcholine is from egg or soyabeans.
 110. The method of claim 79, wherein the surfactant is apolyethyleneglycol-sorbitan-monolaurate, apolyethyleneglycol-sorbitan-monooleate, apolyethyleneglycol-monolaurate, a polyethyleneglycol-monooleate, apolyethyleneglycol-monolaurate-ether, or apolyethyleneglycol-monooleate-ether.
 111. The method of claim 79,wherein the composition further comprises a thickening agent, anantioxidant, or a microbicide.
 112. The method of claim 79, wherein thephosphatidylcholine and the surfactant are present in a molar ratio ofbetween about 40/1 and about 10/1.
 113. The method of claim 79, whereinthe phosphatidylcholine and the surfactant are present in a molar ratioof between about 30/1 and about 7.5/1.
 114. The method of claim 113,wherein the phosphatidylcholine and the surfactant are present in amolar ratio of between about 20/1 and about 10/1.
 115. The method ofclaim 114, wherein the pH is between 0.2 and 2.2 pH units above the pKaof the NSAID.
 116. The method of claim 79, wherein the composition isapplied in a non-occlusive patch.